{"id": "src_201", "tier": 1, "type": "journal", "title": "Novel endo-alpha-N-acetylgalactosaminidases with broader substrate specificity", "authors": ["Koutsioulis D", "Landry D", "Guthrie EP"], "year": 2008, "venue": "Glycobiology", "doi": "10.1093/glycob/cwn069", "pmid": "18635885", "pmcid": "PMC2553423", "url": "https://pubmed.ncbi.nlm.nih.gov/18635885/", "date": "2008-10", "score": 9.0, "authority": 2.5, "recency": 1.0, "primacy": 2.0, "verifiability": 2.0, "coi": 0.5, "conflict_of_interest": "Authors affiliated with NEB Inc", "blacklist_checked": true, "retraction_checked": true, "note": "EngEF (E. faecalis) 序列进入公共领域的核心文献，先公开奠定 FTO 基础；作者为 NEB 雇员但已在学术期刊公开发表", "used_in": ["ch03", "ch12"]}
{"id": "src_202", "tier": 1, "type": "journal", "title": "Molecular cloning, expression, and characterization of a novel endo-alpha-N-acetylgalactosaminidase from Enterococcus faecalis", "authors": ["Goda HM", "Ushigusa K", "Ito H", "Okino N", "Narimatsu H", "Ito M"], "year": 2008, "venue": "Biochem Biophys Res Commun", "doi": "10.1016/j.bbrc.2008.08.065", "pmid": "18725192", "url": "https://www.sciencedirect.com/science/article/abs/pii/S0006291X08015714", "date": "2008-10-31", "score": 8.5, "authority": 1.5, "recency": 1.0, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "note": "日本团队独立克隆 E. faecalis GH101，与 Koutsioulis 2008 同年独立发表，相互印证 EngEF 序列公共领域地位", "used_in": ["ch03", "ch12"]}
{"id": "src_203", "tier": 1, "type": "regulatory", "title": "America Invents Act (AIA), Pub.L. 112-29, 35 U.S.C.", "authors": ["US Congress"], "year": 2011, "venue": "US Federal Law", "doi": null, "url": "https://www.uspto.gov/sites/default/files/aia_implementation/20110916-pub-l112-029.pdf", "date": "2011-09-16", "score": 9.5, "authority": 3.0, "recency": 1.5, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "note": "美国专利法 AIA 先申请制与现有技术规则，第三方公开不享有宽限期豁免", "used_in": ["ch03"]}
{"id": "src_204", "tier": 1, "type": "journal", "title": "The structural basis for T-antigen hydrolysis by Streptococcus pneumoniae: a target for structure-based vaccine design", "authors": ["Caines ME", "Zhu H", "Vuckovic M", "Willis LM", "Withers SG", "Wakarchuk WW", "Strynadka NC"], "year": 2008, "venue": "J Biol Chem", "doi": "10.1074/jbc.C800150200", "pmid": "18784084", "url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC4646209/", "date": "2008-11-14", "score": 9.2, "authority": 2.5, "recency": 1.0, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "note": "SpGH101 首个晶体结构（2.9 Å），明确为学术公开，确立 SpGH101 结构进入公共领域", "used_in": ["ch03"]}
{"id": "src_205", "tier": 1, "type": "journal", "title": "Mechanistic investigation of the endo-alpha-N-acetylgalactosaminidase from Streptococcus pneumoniae R6", "authors": ["Willis LM", "Zhang R", "Reid A", "Withers SG", "Wakarchuk WW"], "year": 2009, "venue": "Biochemistry", "doi": "10.1021/bi9013825", "pmid": "19788271", "url": "https://www.chem.ubc.ca/mechanistic-investigation-endo-alpha-n-acetylgalactosaminidase-streptococcus-pneumoniae-r6", "date": "2009-11-03", "score": 9.0, "authority": 2.5, "recency": 1.0, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "note": "确认 SpGH101 催化残基 D764（亲核）和 E796（广义酸碱），机制研究进入公共领域", "used_in": ["ch03"]}
{"id": "src_206", "tier": 1, "type": "patent", "title": "EP3149034B1 - Deglycosylation reagents and methods", "authors": ["Magnelli P", "Guthrie E", "Taron CH", "Xu MQ", "Buswell J"], "assignee": "New England Biolabs Inc", "year": 2022, "priority_date": "2014-05-30", "filing_date": "2015-05-29", "grant_date": "2022-07-13", "expiry_date": "2035-05-29", "url": "https://patents.google.com/patent/EP3149034B1/en", "date": "2022-07-13", "score": 9.5, "authority": 3.0, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 0.5, "conflict_of_interest": "Assignee NEB is commercial competitor", "blacklist_checked": true, "retraction_checked": false, "note": "NEB 核心脱糖基化专利；独立权利要求聚焦 N-糖苷酶+非 SDS 表面活性剂体系；O-糖苷酶仅在从属权利要求中出现。预计 2035 年到期，中国同族专利状态需 CNIPA 独立核查", "used_in": ["ch03", "ch12"]}
{"id": "src_207", "tier": 1, "type": "journal", "title": "Structural basis of mammalian mucin processing by the human gut O-glycopeptidase OgpA from Akkermansia muciniphila", "authors": ["Trastoy B", "Naegeli A", "Anso I", "Sjogren J", "Guerin ME"], "year": 2020, "venue": "Nat Commun", "doi": "10.1038/s41467-020-18707-y", "url": "https://www.genovis.com/blog/the-ctrystal-structure-of-operator-reveals-o-glycan-substrate-specificity/", "date": "2020-10", "score": 9.0, "authority": 3.0, "recency": 2.0, "primacy": 2.0, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": "Co-author Sjogren affiliated with Genovis AB", "blacklist_checked": true, "retraction_checked": true, "note": "OpeRATOR 晶体结构公开，是下一代酶结构进入公共领域的里程碑；但 Genovis 商业化工艺可能仍受保护", "used_in": ["ch03"]}
{"id": "src_208", "tier": 1, "type": "journal", "title": "Design of a mucin-selective protease for targeted degradation of cancer-associated mucins", "authors": ["Tender GS", "Bertozzi CR", "et al"], "year": 2023, "venue": "Nat Biotechnol", "doi": "10.1038/s41587-023-01840-6", "url": "https://www.nature.com/articles/s41587-023-01840-6", "date": "2023-08-03", "score": 9.5, "authority": 3.0, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 0.5, "conflict_of_interest": "Stanford University holds related patents licensed to Palleon", "blacklist_checked": true, "retraction_checked": true, "note": "eStcE 为黏蛋白选择性蛋白酶（切肽键），与本项目 O-糖苷酶（切糖苷键）属不同酶类，不构成直接 FTO 障碍", "used_in": ["ch03"]}
{"id": "src_209", "tier": 1, "type": "journal", "title": "A high-throughput screening platform for enzymes active on mucin-type O-glycoproteins", "authors": ["Wardman JF", "Sim L", "Liu J", "Howard TA", "Geissner A", "Danby PM", "Boraston AB", "Wakarchuk WW", "Withers SG"], "year": 2023, "venue": "Nat Methods", "doi": "10.1038/s41592-023-01961-7", "pmid": "37592157", "url": "https://pubmed.ncbi.nlm.nih.gov/37592157/", "date": "2023-08-17", "score": 9.5, "authority": 3.0, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 0.5, "conflict_of_interest": "UBC holds patent on this platform methodology", "blacklist_checked": true, "retraction_checked": true, "note": "Withers/UBC 高通量 O-糖肽酶筛选平台（FRET 探针+液滴微流控），已申请 UBC 专利；方法受保护，但筛选所得新酶序列不受此专利覆盖", "used_in": ["ch03"]}
{"id": "src_210", "tier": 1, "type": "journal", "title": "Discovery and Development of Promiscuous O-Glycan Hydrolases for Removal of Intact Sialyl T-Antigen", "authors": ["Wardman JF", "Rahfeld P", "Liu F", "Morgan-Lang C", "Sim L", "Hallam SJ", "Withers SG"], "year": 2021, "venue": "ACS Chem Biol", "doi": "10.1021/acschembio.1c00316", "pmid": "34309358", "url": "https://pubmed.ncbi.nlm.nih.gov/34309358/", "date": "2021-07-26", "score": 9.2, "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 0.5, "conflict_of_interest": "Withers group has filed patents on related technologies (UBC)", "blacklist_checked": true, "retraction_checked": true, "note": "SpGH101 Q868G 突变体首次公开报道，可切除完整唾液酸 T-抗原；宏基因组筛选发现 GH101 家族隐性 STAg 水解活性", "used_in": ["ch03"]}
{"id": "src_211", "tier": 2, "type": "thesis", "title": "Discovery and engineering of enzymes for the manipulation of glycoproteins (PhD Thesis)", "authors": ["Wardman, Jacob Franklin"], "institution": "University of British Columbia", "supervisor": "Withers SG", "year": 2023, "url": "https://open.library.ubc.ca/soa/cIRcle/collections/ubctheses/24/items/1.0438625", "date": "2023", "score": 7.5, "authority": 2.0, "recency": 2.0, "primacy": 1.5, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": "UBC academic thesis, independent research", "blacklist_checked": true, "retraction_checked": false, "note": "系统描述 STAg 水解酶发现、超高通量液滴筛选平台及最优突变体（140倍活性提升）", "used_in": ["ch03"]}
{"id": "src_212", "tier": 1, "type": "journal", "title": "Dual functional POGases from bacteria encompassing broader O-glycanase and adhesin activities", "authors": ["See Nature Communications 2025 author list"], "year": 2025, "venue": "Nat Commun", "doi": "10.1038/s41467-025-57143-8", "pmid": "40000644", "url": "https://www.nature.com/articles/s41467-025-57143-8", "date": "2025-02-25", "score": 9.3, "authority": 3.0, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 0.3, "conflict_of_interest": "Need to check if patent filed before publication", "blacklist_checked": true, "retraction_checked": true, "note": "2025 年最新 GH101 研究，POGase 宽谱酶（sialyl Core 1/2/3）；Kcat/Km 比 EngEF 高 >300 倍；Motif-1/2/3 三段特征序列与已知酶不同，为新进者提供骨架参考。需核查发表前是否有专利申请", "used_in": ["ch03", "ch12"]}
{"id": "src_213", "tier": 1, "type": "journal", "title": "Reshaping of a Glycoside Hydrolase Active Site through Expression in Vivo Directed Evolution", "authors": ["Withers SG group"], "year": 2024, "venue": "ACS Cent Sci", "doi": "10.1021/acscentsci.5c01227", "url": "https://pubs.acs.org/doi/10.1021/acscentsci.5c01227", "date": "2024-2025", "score": 9.0, "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 0.5, "conflict_of_interest": "UBC Withers group, related patents may be filed", "blacklist_checked": true, "retraction_checked": true, "note": "SpGH101 Q868G 定向进化后活性提升 140 倍，证明 Withers/UBC 正在积极推进该方向专利布局，新进者需选择 EngEF 同源位点或全新骨架", "used_in": ["ch03"]}
{"id": "src_101", "tier": 1, "type": "journal", "title": "Identification and Molecular Cloning of a Novel GH Family of Core 1 Type O-Glycan-Specific Endo-alpha-N-Acetylgalactosaminidase from Bifidobacterium longum", "url": "https://pubmed.ncbi.nlm.nih.gov/16141207/", "doi": "10.1074/jbc.M506874200", "year": 2005, "venue": "Journal of Biological Chemistry", "score": 8.5, "authority": 2.5, "recency": 1.0, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "note": "GH101家族创始文献，确认TIM-barrel结构和GH13同源性，保留型机制", "used_in": ["ch02"]}
{"id": "src_102", "tier": 1, "type": "structural", "title": "SpGH101 Crystal Structure (PDB 3ECQ, 2.9A) - Pluvinage/Willis et al.", "url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC2953539/", "doi": "10.1074/jbc.M110.121517", "year": 2010, "venue": "Journal of Biological Chemistry", "score": 9.0, "authority": 3.0, "recency": 1.2, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "note": "SpGH101首个晶体结构，2.9Ang分辨率，PDB 3ECQ", "used_in": ["ch02"]}
{"id": "src_103", "tier": 1, "type": "journal", "title": "Novel endo-alpha-N-acetylgalactosaminidases with broader substrate specificity", "url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC2553423/", "doi": "10.1093/glycob/cwn069", "year": 2008, "venue": "Glycobiology", "score": 8.0, "authority": 2.0, "recency": 1.0, "primacy": 2.0, "verifiability": 2.0, "coi": 0.0, "conflict_of_interest": "NEB资助，作者为NEB员工，已披露", "note": "关键：NEB内部选型报告。EngEF kcat=51.17 s-1 for Core1，最高；Core3 100%；E.coli pET-21a验证", "used_in": ["ch02"], "key_data": {"EngEF_kcat_Core1": "51.17 s-1", "EngEF_Km_Core1": "47.85 uM", "EngSP_kcat_Core1": "10.51 s-1", "expression_host": "E.coli T7 Express lysY/pET-21a"}}
{"id": "src_106", "tier": 2, "type": "database", "title": "BRENDA EC 3.2.1.97 - E. faecalis B5UB72 (EngEF)", "url": "https://www.brenda-enzymes.org/enzyme.php?ecno=3.2.1.97&UniProtAcc=B5UB72&OrganismID=2095", "doi": null, "year": 2026, "venue": "BRENDA (TU Braunschweig)", "score": 7.0, "authority": 2.0, "recency": 2.0, "primacy": 1.5, "verifiability": 1.5, "coi": 1.0, "note": "EngEF动力学参数独立验证来源，与Koutsioulis 2008交叉一致", "used_in": ["ch02"]}
{"id": "src_107", "tier": 2, "type": "product_db", "title": "O-Glycosidase 324716 - Merck Sigma-Aldrich Product Page", "url": "https://www.sigmaaldrich.com/DE/en/product/mm/324716", "doi": null, "year": 2025, "venue": "Merck/Sigma-Aldrich", "score": 6.5, "authority": 2.0, "recency": 2.0, "primacy": 1.5, "verifiability": 1.5, "coi": 0.5, "note": "SpGH101 E.coli重组，比活>=10 units/mg，6项旁活性阴性，适合LC-MS", "used_in": ["ch02"]}
{"id": "src_108", "tier": 2, "type": "technical", "title": "Libios EngEF (E-OGLYEF) Technical Data Sheet, Lot 130201c", "url": "https://libios.fr/imgfr/produit/fichier/1_e-oglyef-data.pdf", "doi": null, "year": 2017, "venue": "Libios", "score": 6.0, "authority": 1.5, "recency": 0.5, "primacy": 1.5, "verifiability": 1.5, "coi": 0.5, "note": "EngEF批次实测数据：MW 158800 Da，pI 5.2，比活3.0 U/mg，pH 7.5", "used_in": ["ch02"]}
{"id": "src_109", "tier": 2, "type": "product_db", "title": "NEB O-Glycosidase P0733 Product Page + NEB Glycoproteomics Technical Guide", "url": "https://www.neb.com/en-us/products/p0733-o-glycosidase", "doi": null, "year": 2025, "venue": "New England Biolabs", "score": 6.0, "authority": 2.0, "recency": 2.0, "primacy": 1.5, "verifiability": 1.0, "coi": 0.0, "note": "P0733 40M units/mL，GlycoBuffer 2，37°C，需Neuraminidase预处理", "used_in": ["ch02", "ch12"]}
{"id": "src_110", "tier": 1, "type": "journal", "title": "Discovery and Development of Promiscuous O-Glycan Hydrolases (Q868G SpGH101 mutant)", "url": "https://pubs.acs.org/doi/10.1021/acschembio.1c00316", "doi": "10.1021/acschembio.1c00316", "year": 2021, "venue": "ACS Chemical Biology", "score": 9.0, "authority": 2.5, "recency": 1.5, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "note": "Wardman/Withers Q868G单点突变扩展底物谱，kcat/Km对sialyl T-antigen提升4.8倍", "used_in": ["ch02"], "key_data": {"mutation": "Q868G", "improvement_MU_STAg": "4.8x"}}
{"id": "src_111", "tier": 2, "type": "conference", "title": "Novel Enzymes for O-glycan Analysis - Genovis CASSS 2018 Poster", "url": "https://www.genovis.com/wp-content/uploads/2018-CASSS-Novel-Enzymes-for-O-glycan-Analysis.pdf", "doi": null, "year": 2018, "venue": "CASSS/Genovis", "score": 5.5, "authority": 1.5, "recency": 1.0, "primacy": 1.0, "verifiability": 1.0, "coi": 0.0, "conflict_of_interest": "Genovis商业推广材料", "note": "独立验证EngEF唾液酸依赖局限；S.oralis O-糖苷酶优于E.faecalis EngEF", "used_in": ["ch02"]}
{"id": "src_112", "tier": 1, "type": "journal", "title": "Dual functional POGases from bacteria encompassing broader O-glycanase and adhesin activities", "url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC11861894/", "doi": "10.1038/s41467-025-57143-8", "year": 2025, "venue": "Nature Communications", "score": 9.0, "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "note": "POGase宽谱酶发现；EngCP天然G868仍无sialyl Core 1活性，是Q868G反方证据。EngEF Km Core1=15.3 uM", "used_in": ["ch02"]}
{"id": "src_113", "tier": 2, "type": "product_db", "title": "SHuffle T7 Competent E. coli C3026 - NEB Product Page", "url": "https://www.neb.com/en-us/products/c3026-shuffle-t7-competent-e-coli", "doi": null, "year": 2025, "venue": "New England Biolabs", "score": 6.5, "authority": 2.0, "recency": 2.0, "primacy": 1.5, "verifiability": 1.5, "coi": 0.5, "note": "SHuffle T7：Δgor ΔtrxB胞质氧化，DsbC组成型表达，BSL-1，适合复杂蛋白表达", "used_in": ["ch02"]}
{"id": "src_114", "tier": 1, "type": "journal", "title": "E. coli Cytoplasmic Expression of Disulfide-Bonded Proteins: SHuffle vs CyDisCo Side-by-Side Comparison", "url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC11180911/", "doi": "10.3390/ijms25115836", "year": 2024, "venue": "IJMS (MDPI)", "score": 7.5, "authority": 2.0, "recency": 2.0, "primacy": 2.0, "verifiability": 1.5, "coi": 1.0, "note": "SHuffle在T7富培养基下产量5-450 mg/L；CyDisCo在9/10蛋白更高产量（反方证据）", "used_in": ["ch02"]}
{"id": "src_115", "tier": 2, "type": "product_db", "title": "NEB Endo S2 P0761 QC Specifications - 类比参照", "url": "https://www.neb.com/en-us/products/p0761-endo-s2", "doi": null, "year": 2025, "venue": "New England Biolabs", "score": 6.0, "authority": 2.0, "recency": 2.0, "primacy": 1.0, "verifiability": 1.0, "coi": 0.5, "note": "[类比] QC复杂度估算参照，非P0733直接QC规格单", "used_in": ["ch02"]}
{"id": "src_116", "tier": 1, "type": "regulatory", "title": "PHAC PSDS: Enterococcus faecalis and Enterococcus faecium", "url": "https://www.canada.ca/en/public-health/services/laboratory-biosafety-biosecurity/pathogen-safety-data-sheets-risk-assessment/enterococcus-faecalis.html", "doi": null, "year": 2023, "venue": "Public Health Agency of Canada", "score": 9.5, "authority": 3.0, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "note": "官方PSDS，E. faecalis Risk Group 2，Containment Level 2要求", "used_in": ["ch02"]}
{"id": "src_117", "tier": 2, "type": "regulatory", "title": "Samuel Merritt University Biosafety Manual Ver 2.0", "url": "https://www.samuelmerritt.edu/sites/default/files/2021-07/Biosafety%20Manual%20Ver%202.0%202021_1.pdf", "doi": null, "year": 2021, "venue": "Samuel Merritt University EHS", "score": 7.0, "authority": 2.0, "recency": 1.5, "primacy": 1.5, "verifiability": 1.5, "coi": 1.0, "note": "明确E. faecalis需BSL-2实验室", "used_in": ["ch02"]}
{"id": "src_118", "tier": 2, "type": "regulatory", "title": "E. faecalis & E. faecium Agent Safety Guide - USC EHS", "url": "https://sc.edu/about/offices_and_divisions/ehs/research_and_laboratory_safety/biological_safety/biological_agent_safety_guides/agent_safety_guides/e-faecalis-and-e-faecium-safety-guide.pdf", "doi": null, "year": 2023, "venue": "USC EHS", "score": 7.0, "authority": 2.0, "recency": 2.0, "primacy": 1.5, "verifiability": 1.5, "coi": 1.0, "note": "BSL-2实验室要求，E.coli表达路径可绕开BSL-2", "used_in": ["ch02"]}
{"id": "src_119", "tier": 2, "type": "company", "title": "GenScript BacPower E. coli Platform / Bestzyme CDMO Capability", "url": "https://www.genscript.com/protein-expression-services.html", "doi": null, "year": 2025, "venue": "GenScript/Bestzyme", "score": 5.5, "authority": 1.5, "recency": 2.0, "primacy": 1.0, "verifiability": 1.0, "coi": 0.0, "note": "[待验证] 金斯瑞BacPower 15g/L E.coli，2000L发酵规模；百斯杰2023年融资2.5亿RMB", "used_in": ["ch02"]}
{"id": "src_120", "tier": 2, "type": "journal", "title": "Using Glycosidases to Remove, Trim, or Modify Glycans on Therapeutic Proteins", "url": "https://www.bioprocessintl.com/cell-line-development/using-glycosidases-to-remove-trim-or-modify-glycans-on-therapeutic-proteins", "doi": null, "year": 2022, "venue": "BioProcess International", "score": 6.5, "authority": 2.0, "recency": 1.5, "primacy": 1.5, "verifiability": 1.5, "coi": 0.5, "note": "综述指出无已知广谱O-糖苷酶可切所有O-糖链，两款商业酶底物局限性明确（反方证据）", "used_in": ["ch02"]}
{"id": "src_301", "tier": 2, "type": "journal", "title": "Optimizing recombinant protein expression via automated induction profiling in microtiter plates at different temperatures", "authors": ["Sivashanmugam A", "Murray V", "Cui C", "Zhang Y", "Wang J", "Li Q"], "year": 2017, "venue": "BMC Biotechnology (approx.)/PMC5706349", "doi": null, "pmcid": "PMC5706349", "url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC5706349/", "accessed_at": "2026-04-21", "score": 7.5, "authority": 2.0, "recency": 1.5, "primacy": 2.0, "verifiability": 1.5, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "note": "系统评估不同温度下IPTG浓度优化：最优IPTG为0.05–0.1 mM；高温时最优IPTG更低；支持低浓度IPTG用于改善可溶性", "used_in": ["ch04"]}
{"id": "src_302", "tier": 1, "type": "journal", "title": "Production of soluble eukaryotic recombinant proteins in E. coli is favoured in early log-phase cultures induced at low temperature", "authors": ["San-Miguel T", "Pérez-Bermejo P", "Gavilanes F"], "year": 2013, "venue": "SpringerPlus", "doi": "10.1186/2193-1801-2-89", "pmcid": "PMC3602615", "url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC3602615/", "accessed_at": "2026-04-21", "score": 8.0, "authority": 2.0, "recency": 1.5, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "note": "系统证明低温（16–23°C）诱导显著改善>80 kDa蛋白可溶性；早期对数期+低温组合最优；支持EngEF低温诱导策略", "used_in": ["ch04", "ch12"]}
{"id": "src_303", "tier": 1, "type": "journal", "title": "SHuffle, a novel Escherichia coli protein expression strain capable of correctly folding disulfide bonded proteins in its cytoplasm", "authors": ["Lobstein J", "Emrich CA", "Jeans C", "Faulkner M", "Riggs P", "Berkmen M"], "year": 2012, "venue": "Microbial Cell Factories", "doi": "10.1186/1475-2859-11-56", "pmcid": "PMC3526497", "url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC3526497/", "accessed_at": "2026-04-21", "score": 8.5, "authority": 2.5, "recency": 1.5, "primacy": 2.0, "verifiability": 2.0, "coi": 0.5, "conflict_of_interest": "First author affiliated with NEB (SHuffle developer)", "blacklist_checked": true, "retraction_checked": true, "note": "SHuffle T7菌株原始描述论文：Δgor ΔtrxB氧化胞质+染色体DsbC；T7富培养基下5–450 mg/L产量区间；适合多二硫键蛋白", "used_in": ["ch04", "ch12"]}
{"id": "src_304", "tier": 1, "type": "journal", "title": "The Ability to Enhance the Solubility of Its Fusion Partners Is an Intrinsic Property of Maltose-Binding Protein but Their Folding Is Either Spontaneous or Chaperone-Mediated", "authors": ["Fox JD", "Waugh DS"], "year": 2012, "venue": "PLOS ONE", "doi": "10.1371/journal.pone.0049589", "url": "https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0049589", "accessed_at": "2026-04-21", "score": 8.0, "authority": 2.5, "recency": 1.5, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "note": "MBP溶解性增强机制（holdase）的系统研究；MBP融合将多种蛋白可溶性从<5%提升至30–60%；与NusA对比；作为标准溶解性标签被广泛引用", "used_in": ["ch04", "ch12"]}
{"id": "src_305", "tier": 1, "type": "journal", "title": "Enhancing the solubility of recombinant proteins in Escherichia coli by using hexahistidine-tagged maltose-binding protein as a fusion partner", "authors": ["Waugh DS"], "year": 2011, "venue": "Methods Mol Biol", "doi": null, "pmid": "21125392", "url": "https://pubmed.ncbi.nlm.nih.gov/21125392/", "accessed_at": "2026-04-21", "score": 7.5, "authority": 2.0, "recency": 1.5, "primacy": 2.0, "verifiability": 1.5, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "note": "His6-MBP双功能标签方法综述：允许IMAC一步纯化；MBP为最有效溶解性增强标签之一；可与任何目标蛋白融合", "used_in": ["ch04"]}
{"id": "src_306", "tier": 2, "type": "journal", "title": "Fusion tags for protein solubility, purification and immunogenicity in Escherichia coli: the novel Fh8 system", "authors": ["Costa SJ", "Almeida A", "Castro A", "Domingues L"], "year": 2014, "venue": "Frontiers in Microbiology", "doi": "10.3389/fmicb.2014.00063", "url": "https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2014.00063/full", "accessed_at": "2026-04-21", "score": 7.0, "authority": 2.0, "recency": 1.5, "primacy": 1.5, "verifiability": 1.5, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "note": "MBP（43 kDa）和SUMO（~12 kDa）融合标签综述；TEV蛋白酶切除方案；SUMO适合N端敏感应用；支持MBP/SUMO策略选择依据", "used_in": ["ch04"]}
{"id": "src_307", "tier": 1, "type": "journal", "title": "Engineering Bacillus subtilis for high-value bioproduction: recent advances and applications", "authors": ["PMC12341298 author list"], "year": 2025, "venue": "Microbial Cell Factories (PMC12341298)", "doi": null, "pmcid": "PMC12341298", "url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC12341298/", "accessed_at": "2026-04-21", "score": 8.5, "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "note": "B. subtilis占全球工业酶市场~60%；GRAS地位；天然分泌能力优势；WB600菌株分泌天冬酰胺酶407.6 U/mL；对比E. coli的包涵体劣势", "used_in": ["ch04", "ch12"]}
{"id": "src_308", "tier": 2, "type": "journal", "title": "Recombinant protein secretion by Bacillus subtilis and Lactococcus lactis: pathways, applications, and innovation potential", "authors": ["van Dijl JM", "Hecker M"], "year": 2021, "venue": "Applied Microbiology and Biotechnology (PMC8314018)", "doi": null, "pmcid": "PMC8314018", "url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC8314018/", "accessed_at": "2026-04-21", "score": 8.0, "authority": 2.5, "recency": 2.0, "primacy": 1.5, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "note": "欧洲工业酶年营业额>20亿欧元；洗涤剂蛋白酶年产900吨纯酶；B. subtilis分泌能力综述；Sec通路详细机制", "used_in": ["ch04"]}
{"id": "src_309", "tier": 1, "type": "regulatory", "title": "Bacillus subtilis NRRL 68054 GRAS Notice - FDA GRN Database", "authors": ["FDA"], "year": 2025, "venue": "US FDA GRAS Notice Inventory", "doi": null, "url": "https://downloads.regulations.gov/FDA-2025-N-1927-0061/attachment_1.pdf", "accessed_at": "2026-04-21", "score": 9.0, "authority": 3.0, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "note": "FDA GRN库中有25条B. subtilis相关GRAS通知，20条已获no-questions回函；证明B. subtilis GRAS地位的一级监管文件", "used_in": ["ch04"]}
{"id": "src_310", "tier": 1, "type": "journal", "title": "Bottleneck in secretion of alpha-amylase in Bacillus subtilis", "authors": ["Krishnappa L et al."], "year": 2017, "venue": "Microbial Cell Factories", "doi": "10.1186/s12934-017-0739-5", "pmid": "28724440", "url": "https://pubmed.ncbi.nlm.nih.gov/28724440/", "accessed_at": "2026-04-21", "score": 8.0, "authority": 2.5, "recency": 1.5, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "note": "B. subtilis Sec通路分泌>80 kDa大蛋白存在瓶颈（反方证据）；α-淀粉酶案例；提示EngEF（108 kDa）迁移B. subtilis需截短体策略", "used_in": ["ch04"]}
{"id": "src_311", "tier": 1, "type": "journal", "title": "Pichia pastoris: A highly successful expression system for optimal synthesis of foreign proteins", "authors": ["Ahmad M et al."], "year": 2014, "venue": "Molecular Biotechnology (PMC7228273)", "doi": null, "pmcid": "PMC7228273", "url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC7228273/", "accessed_at": "2026-04-21", "score": 7.5, "authority": 2.0, "recency": 1.5, "primacy": 1.5, "verifiability": 1.5, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "note": "Pichia OCH1控制超糖基化；O-甘露糖化对酶活性的干扰综述；GlycoSwitch技术局限（主要针对N-糖，O-甘露糖化方案不完善）；支持否决Pichia用于O-糖苷酶的关键综述", "used_in": ["ch04", "ch12"]}
{"id": "src_312", "tier": 1, "type": "journal", "title": "Identification and Functional Characterization of Glycosylation of Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris", "authors": ["Li et al."], "year": 2015, "venue": "PLOS ONE (PMC4689512)", "doi": null, "pmcid": "PMC4689512", "url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC4689512/", "accessed_at": "2026-04-21", "score": 7.5, "authority": 2.0, "recency": 1.5, "primacy": 2.0, "verifiability": 1.5, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "note": "Pichia中PDGF-BB的O-甘露糖化定位（Thr96/Thr104）及其对功能影响；O-甘露糖化异质性导致蛋白结构/功能改变的案例证据", "used_in": ["ch04"]}
{"id": "src_313", "tier": 1, "type": "journal", "title": "Engineering complex-type N-glycosylation in Pichia pastoris using GlycoSwitch technology", "authors": ["Jacobs PP et al."], "year": 2009, "venue": "Nature Protocols", "doi": "10.1038/nprot.2008.213", "pmid": "19131957", "url": "https://pubmed.ncbi.nlm.nih.gov/19131957/", "accessed_at": "2026-04-21", "score": 8.5, "authority": 2.5, "recency": 1.0, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "note": "GlycoSwitch技术：OCH1敲除+异源糖基转移酶改造Pichia N-糖基化；每步工程约需3周；注意该技术主要针对N-糖，O-甘露糖化问题未解决（反方/限制证据）", "used_in": ["ch04", "ch12"]}
{"id": "src_314", "tier": 1, "type": "regulatory", "title": "CDC Quick Learn: Recognize the four Biosafety Levels", "authors": ["US CDC"], "year": 2024, "venue": "US Centers for Disease Control and Prevention", "doi": null, "url": "https://www.cdc.gov/training/quicklearns/biosafety/", "accessed_at": "2026-04-21", "score": 9.5, "authority": 3.0, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "note": "BSL-2要求：BSC操作+门禁+高压灭菌废物+工作人员培训。E. coli K-12衍生株（BL21/SHuffle）为BSL-1，全程兼容普通CDMO。来源CDC官方培训材料", "used_in": ["ch04"]}
{"id": "src_502", "tier": 2, "score": 6.5, "type": "product_catalog", "title": "O-Glycosidase from Streptococcus pneumoniae G1163 — Merck Sigma-Aldrich via SLS UK", "authors": ["Merck Sigma-Aldrich / Scientific Laboratory Supplies"], "year": 2025, "venue": "scientificlabs.com (Merck authorized UK distributor)", "url": "https://www.scientificlabs.com/en/product/protein-assays/g1163-.04un", "accessed_at": "2026-04-20", "authority": 2.0, "recency": 2.0, "primacy": 1.0, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"G1163_price_EUR": 621.68, "region": "UK/Ireland only", "concentration": ">=800 units/mL", "form": "buffered aqueous solution", "mol_wt": "180 kDa"}, "used_in": ["ch06"], "notes": "英国区报价；美国区 Sigma-Aldrich G1163 定价需登录后查看，无法获取，本信源仅作欧洲市场参照"}
{"id": "src_503", "tier": 1, "score": 8.8, "type": "financial", "title": "Bio-Techne Corporation (TECH) Income Statement FY2021-FY2025 — SEC 10-K Data", "authors": ["Bio-Techne Corporation", "StockAnalysis.com (Fiscal.ai data aggregator)"], "year": 2025, "venue": "SEC 10-K / StockAnalysis", "url": "https://stockanalysis.com/stocks/tech/financials/", "accessed_at": "2026-04-20", "authority": 3.0, "recency": 2.0, "primacy": 2.0, "verifiability": 1.5, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"FY2025_gross_margin": "64.80%", "FY2024_gross_margin": "66.41%", "FY2023_gross_margin": "67.72%", "FY2022_gross_margin": "68.42%", "FY2021_gross_margin": "67.97%", "5yr_avg_gross_margin": "~67%", "FY2025_revenue_M_USD": 1220}, "used_in": ["ch06"], "notes": "Bio-Techne 作为生命科学试剂行业上市公司参照；NEB 毛利率类比估算基准；FY 截止 6 月 30 日"}
{"id": "src_504", "tier": 3, "score": 4.0, "type": "internal_estimate", "title": "重组酶 E.coli 表达体系通用成本结构假设（内部推算）", "authors": ["dr-analyst"], "year": 2026, "venue": "本研究内部逻辑推导", "url": null, "accessed_at": "2026-04-20", "authority": 0.0, "recency": 2.0, "primacy": 0.5, "verifiability": 0.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": false, "retraction_checked": false, "key_data": {"assumption": "国产重组酶与 NEB 同等 E.coli 表达体系，可类比 Bio-Techne 毛利结构"}, "used_in": ["ch06"], "notes": "仅作估算支撑，不作为结论唯一来源"}
{"id": "src_505", "tier": 2, "score": 6.8, "type": "equity_research", "title": "打破生物试剂进口垄断，国产细分龙头崛起（诺唯赞/翌圣生物深度研报）", "authors": ["国元证券研究所", "东北证券"], "year": 2023, "venue": "Wind/东方财富研报平台", "url": "https://pdf.dfcfw.com/pdf/H3_AP202107151503924611_1.pdf", "accessed_at": "2026-04-20", "authority": 1.5, "recency": 1.5, "primacy": 1.0, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": "券商研报，存在潜在利益关系但数据来源于公开招股书", "blacklist_checked": true, "retraction_checked": false, "key_data": {"NEB_China_share_2020": "9.3%", "Yisheng_domestic_share": "24.2%", "Vazyme_domestic_share": "12.1%", "new_entrant_first_year_share": "<5%（类比）"}, "used_in": ["ch06"], "notes": "数据主要针对分子酶品类（PCR 酶、逆转录酶），与 O-糖苷酶品类存在差异，外推需谨慎"}
{"id": "src_506", "tier": 2, "score": 8.0, "type": "consulting_report", "title": "China's Life Sciences Sector in Transition: How Suppliers Can Thrive in the Next Chapter", "authors": ["L.E.K. Consulting"], "year": 2024, "venue": "L.E.K. Consulting Insights", "url": "https://www.lek.com/insights/hea/cn/ei/chinas-life-sciences-sector-transition-how-suppliers-can-thrive-next-chapter", "accessed_at": "2026-04-20", "authority": 2.5, "recency": 2.0, "primacy": 1.5, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"finding_1": "中国市场竞争加剧使 MNC 毛利受压", "finding_2": "价格竞争在商品化品类最为激烈", "finding_3": "高端细分 MNC 仍保持质量优势", "finding_4": "中国替代在不同细分差异显著"}, "used_in": ["ch06", "ch12"], "notes": "L.E.K. 是 Tier 2 咨询机构，2024 年报告，时效性高，作为反方证据使用"}
{"id": "src_507", "tier": 1, "score": 8.5, "type": "product_catalog_and_annual_report", "title": "Genovis OpeRATOR Lyophilized Product Page + Year-End Report January-December 2025", "authors": ["Genovis AB"], "year": 2025, "venue": "Genovis Official Website / Genovis Investor Relations", "url_product": "https://www.genovis.com/product-group/operator/", "url_ar": "https://investor.genovis.com/en/mfn_news/genovis-ab-year-end-report-january-december-2025/", "accessed_at": "2026-04-20", "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"OpeRATOR_price_EUR": 1251.0, "OpeRATOR_catalog": "G2-OP1-020", "OpeRATOR_size": "2000 units lyophilized", "FY2025_net_sales_SEK_thousands": 128946, "FY2025_EBITDA_SEK_thousands": 33558, "FY2025_EBITDA_margin": "~26%", "Q4_2025_EBITDA_margin": "29%", "enzyme_growth_FY2025": "17% (23% FX-adjusted)"}, "used_in": ["ch06", "ch12"], "notes": "OpeRATOR 价格 2026-04-20 查询确认；Genovis 年收入约 SEK 1.29 亿（~€1,110 万），中型酶企业"}
{"id": "src_508", "tier": 1, "score": 8.5, "type": "product_catalog", "title": "O-Glycoprotease (IMPa) P0761S — NEB Product Page", "authors": ["New England Biolabs"], "year": 2026, "venue": "NEB Official Website", "url": "https://www.neb.com/en-us/products/p0761-o-glycoprotease", "accessed_at": "2026-04-20", "authority": 3.0, "recency": 2.0, "primacy": 2.0, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"catalog": "P0761S", "size": "1 x 200 reactions", "concentration": "1,000 units/ml", "source_organism": "Pseudomonas aeruginosa", "price": "未获取（需登录 NEB 账户）"}, "used_in": ["ch06"], "notes": "IMPa 为 NEB 旗下下一代 O-糖蛋白酶，与 Genovis OpeRATOR 构成工程酶市场双寡头；具体售价未获取，需进一步查询"}
{"id": "src_401", "tier": 2, "score": 7.5, "type": "market_report", "title": "Glycomics Market Size, Share, Growth & Trends Report 2030 — Drug Discovery Segment", "authors": ["Mordor Intelligence"], "year": 2025, "venue": "Mordor Intelligence", "url": "https://www.mordorintelligence.com/industry-reports/glycomics-market", "accessed_at": "2026-04-21", "authority": 2.0, "recency": 2.0, "primacy": 1.5, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"pharma_share_2025": "48.62%", "vaccine_cagr": "14.21%", "hospital_cagr": "14.98%", "drug_discovery_share_2025": "45.57%"}, "used_in": ["ch05"], "notes": "制药/生物技术公司2025年贡献48.62%的糖组学市场营收；ADC应用是关键增长驱动"}
{"id": "src_402", "tier": 2, "score": 7.0, "type": "market_report", "title": "Glycobiology Market — Enzyme & Product Analysis (Coherent Market Insights)", "authors": ["Coherent Market Insights"], "year": 2024, "venue": "Coherent Market Insights", "url": "https://www.coherentmarketinsights.com/market-insight/glycobiology-market-3639", "accessed_at": "2026-04-21", "authority": 2.0, "recency": 2.0, "primacy": 1.0, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"enzyme_utilization_share": "40-55%", "asia_pacific_cagr": ">16% (2025-2033)"}, "used_in": ["ch05"], "notes": "酶类产品占糖生物学产品价值约40-55%，是最大品类"}
{"id": "src_403", "tier": 2, "score": 7.5, "type": "market_report", "title": "U.S. Glycomics Market Size & Share | Industry Report, 2030 (Grand View Research)", "authors": ["Grand View Research"], "year": 2024, "venue": "Grand View Research", "url": "https://www.grandviewresearch.com/industry-analysis/us-glycomics-market-report", "accessed_at": "2026-04-21", "authority": 2.0, "recency": 2.0, "primacy": 1.5, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"academic_share_2023": "39%", "drug_discovery_dominant_application": "true"}, "used_in": ["ch05"], "notes": "学术与科研机构2023年占糖组学终端用户约39%，居最高；药物发现占最大应用份额"}
{"id": "src_404", "tier": 1, "score": 9.0, "type": "ipo_prospectus", "title": "翌圣生物科技（上海）股份有限公司招股说明书（科创板上会稿，含Frost & Sullivan中国生物试剂市场数据）", "authors": ["翌圣生物科技（上海）股份有限公司"], "year": 2023, "venue": "上海证券交易所科创板", "url": "http://static.sse.com.cn/stock/disclosure/announcement/c/202306/001182_20230630_JIKV.pdf", "accessed_at": "2026-04-21", "authority": 3.0, "recency": 1.5, "primacy": 2.0, "verifiability": 2.0, "coi": 0.5, "conflict_of_interest": "上市申请文件，公司自身利益相关，但数据引自独立Frost & Sullivan报告", "blacklist_checked": true, "retraction_checked": false, "key_data": {"china_bio_reagent_market_2021_bn_rmb": 183, "import_share_research_institute": "90%", "domestic_share_2021": "10%", "china_bio_reagent_2024_bn_rmb": 258.4, "cagr_2019_2024": "13.8%"}, "used_in": ["ch05", "ch12"], "notes": "2021年中国生物试剂科研市场规模183亿元；进口品牌占科研机构用户市场约90%；是中国生物试剂市场最权威的一手披露来源之一"}
{"id": "src_405", "tier": 1, "score": 9.5, "type": "regulatory_guideline", "title": "ICH Q2(R2) Guideline on Validation of Analytical Procedures — Step 5 (EMA effective June 2024)", "authors": ["ICH", "EMA"], "year": 2024, "venue": "EMA Official Publication", "url": "https://www.ema.europa.eu/en/ich-q2r2-validation-analytical-procedures-scientific-guideline", "doi": null, "accessed_at": "2026-04-21", "authority": 3.0, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"effective_date_ema": "2024-06-14", "key_provision": "试剂变更须评估是否需部分或全部重验证", "revalidation_parameters": ["specificity", "precision", "accuracy", "linearity", "robustness"]}, "used_in": ["ch05", "ch12"], "notes": "ICH Q2(R2) 2023年修订版，EMA 2024年6月生效；引入分析生命周期管理框架；CMC方法验证的最高权威指引"}
{"id": "src_406", "tier": 2, "score": 7.0, "type": "review_meta", "title": "IgA Nephropathy in China: Epidemiology, Clinical Features and Disease Burden (多篇系统综述合并引用)", "authors": ["Zhang et al.; CNKI综述"], "year": 2022, "venue": "PubMed / CNKI 综合", "url": "https://pubmed.ncbi.nlm.nih.gov/?term=IgA+nephropathy+China+prevalence+epidemiology", "accessed_at": "2026-04-21", "authority": 2.0, "recency": 1.5, "primacy": 1.5, "verifiability": 1.5, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"china_IgAN_patients_estimate": "~500万", "global_largest_patient_population": true}, "used_in": ["ch05"], "notes": "[待验证] 500万例数字见于多篇meta分析，建议回溯CNKI原始流行病学调查文献校正"}
{"id": "src_407", "tier": 1, "score": 9.5, "type": "annual_report", "title": "WuXi XDC Cayman Inc. Annual Report 2024", "authors": ["药明合联（WuXi XDC）", "港交所"], "year": 2025, "venue": "港交所上市公司年报（2268.HK）", "url": "https://www.hkexnews.hk/listedco/listconews/sehk/2025/0429/2025042903387.pdf", "accessed_at": "2026-04-21", "authority": 3.0, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 0.5, "conflict_of_interest": "公司自身年报，数据可信但存在选择性披露可能", "blacklist_checked": true, "retraction_checked": false, "key_data": {"iCMC_projects_2024_end": 194, "phase2_plus_projects": 69, "PPQ_commercial_projects": 8, "revenue_2024_bn_rmb": 4.052, "revenue_growth_yoy": "90.8%", "customers_2024": 499, "cumulative_projects_discovery_to_CMC": 45}, "used_in": ["ch05", "ch12"], "notes": "截至2024年12月31日194个正在进行的iCMC整合项目；分析表征使用HPLC/ELISA/LC-MS测量包括糖基化在内的关键参数"}
{"id": "src_408", "tier": 2, "score": 7.0, "type": "investment_report", "title": "How are ADC drug partnerships changing China's biotech landscape? (Invesco, October 2024)", "authors": ["Chris Liu", "Invesco Asset Management"], "year": 2024, "venue": "Invesco APAC Insights", "url": "https://www.invesco.com/content/dam/invesco/apac/en/pdf/insights/2024/october/invesco-how-are-adc-drug-partnerships-changing-chinas-biotech-landscape-oct-2024.pdf", "accessed_at": "2026-04-21", "authority": 2.0, "recency": 2.0, "primacy": 1.0, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": "投资机构，可能有立场偏向中国biotech积极叙事", "blacklist_checked": true, "retraction_checked": false, "key_data": {"global_ADC_clinical_candidates_2024": ">230", "china_ADC_license_out_deals_2022_2024": 35, "us_ADC_license_out_same_period": 25, "china_ADC_market_share_global": "significant and growing"}, "used_in": ["ch05"], "notes": "中国ADC对外授权交易2022-2024年35笔，超过美国25笔，是全球最大授权国；截至2024年全球230+款ADC在临床"}
{"id": "src_409", "tier": 3, "score": 6.0, "type": "equity_research", "title": "华创证券·医药投资观点周周谈第70期：造影剂市场国产替代东风已至（含生命科学服务进口替代板块）", "authors": ["华创证券研究所"], "year": 2024, "venue": "华创证券", "url": "https://aigc.idigital.com.cn（华创证券研报平台）", "accessed_at": "2026-04-21", "authority": 1.5, "recency": 2.0, "primacy": 1.0, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": "券商研报，可能存在推荐标的利益", "blacklist_checked": true, "retraction_checked": false, "key_data": {"bio_reagent_market_2021_bn_rmb": 392, "domestic_share_science_2021": "10%", "domestic_share_molecular_2020": "30%"}, "used_in": ["ch05"], "notes": "仅作辅助引用；核心数据引自公开招股书；生物科研试剂2021年国产份额约10%（科研端）"}
{"id": "src_410", "tier": 1, "score": 9.5, "type": "regulatory_guideline", "title": "Analytical Procedures and Methods Validation for Drugs and Biologics — FDA Guidance for Industry", "authors": ["US FDA"], "year": 2015, "venue": "FDA Guidance Document（现行有效）", "url": "https://www.fda.gov/files/drugs/published/Analytical-Procedures-and-Methods-Validation-for-Drugs-and-Biologics.pdf", "accessed_at": "2026-04-21", "authority": 3.0, "recency": 1.5, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"revalidation_triggers": ["试剂更换", "实验室迁移", "制造工艺变更"], "prior_approval_supplement": "已批准BLA/NDA中分析方法的重大变更需PAS", "regulatory_ref": "21 CFR 601.2 for BLA"}, "used_in": ["ch05"], "notes": "FDA现行有效指引；试剂替换触发再验证或等效性研究；商业化阶段变更需PAS并等待监管审批"}
{"id": "src_411", "tier": 2, "score": 7.5, "type": "industry_guidance", "title": "Best practices for the development, validation and registration of analytical procedures — Implementation of ICH Q2(R2) and Q14 for biologics (BioPhorum)", "authors": ["BioPhorum"], "year": 2025, "venue": "BioPhorum (Pharma industry consortium)", "url": "https://www.biophorum.com/download/best-practices-for-the-development-validation-and-registration-of-analytical-procedures-implementation-of-ich-q2-r2-and-q14-for-biologics/", "accessed_at": "2026-04-21", "authority": 2.5, "recency": 2.0, "primacy": 1.5, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"APLCM_document": "Analytical Procedure Lifecycle Management Document", "APD_in_registration": "Analytical Procedure Document included in regulatory submissions", "change_control_trigger": "Reagent supplier change triggers Change Control under ICH Q2(R2)/Q14 lifecycle framework"}, "used_in": ["ch05"], "notes": "BioPhorum是制药行业联盟（无商业利益冲突）；ICH Q2(R2)+Q14联合实施最佳实践；分析程序档案（APD）中供应商信息一旦注册即形成变更控制壁垒"}
{"id": "src_412", "tier": 1, "score": 8.5, "type": "interim_report", "title": "Genovis AB Interim Report January–September 2024", "authors": ["Genovis AB"], "year": 2024, "venue": "Genovis AB (Nasdaq First North Growth Market)", "url": "https://investor.genovis.com/en/mfn_news/interim-report-january-september-2024/", "accessed_at": "2026-04-21", "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 0.5, "conflict_of_interest": "公司自身财务报告，存在选择性披露可能", "blacklist_checked": true, "retraction_checked": false, "key_data": {"Q3_2024_analytics_enzyme_sales_SEK_thousands": 31600, "Q3_yoy_growth": "24%", "Q3_new_record": true, "growth_driver": "ADC technology large orders", "9M_2024_enzyme_business_growth_fx_adj": "12%"}, "used_in": ["ch05"], "notes": "2024年Q3酶类（analytics）销售额SEK 3,160万，同比增长24%，创单季历史新高；增长主要来自ADC技术相关大单；证明ADC驱动的糖分析酶市场实质性扩张"}
{"id": "src_413", "tier": 2, "score": 8.0, "type": "price_list", "title": "New England Biolabs TCEFS 2025 Pricing List (effective 2025-01-29)", "authors": ["New England Biolabs", "UMass Medical School TCEFS Contract"], "year": 2025, "venue": "UMass Medical School Enzyme Freezer / NEB", "url": "https://www.umassmed.edu/globalassets/enzyme-freezer/documents/neb-tcefs-2025-pricing-list-01-29-25.pdf", "accessed_at": "2026-04-21", "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"P0733S_TCEFS_USD": 137, "P0733L_TCEFS_USD": 525, "E0540S_TCEFS_USD": 190, "P0720S_TCEFS_USD": 87, "P0761S_TCEFS_USD": "not listed", "effective_date": "2025-01-29"}, "used_in": ["ch07", "ch12"], "notes": "NEB TCEFS 2025协议价格表：P0733S $137, P0733L $525, E0540S $190（O-糖苷酶+神经氨酸酶捆绑），P0720S神经氨酸酶$87。这是 NEB O-糖苷酶产品线精确定价的最可靠公开来源"}
{"id": "src_414", "tier": 2, "score": 6.5, "type": "distributor_catalog", "title": "O-Glycosidase & α2-3,6,8 Neuraminidase Bundle E0540S — BIOKÉ NL Product Page", "authors": ["BIOKÉ (NEB Authorized Distributor)"], "year": 2026, "venue": "BIOKÉ B.V. (Netherlands)", "url": "https://www.bioke.com/webshop/neb/e0540.html", "accessed_at": "2026-04-21", "authority": 2.0, "recency": 2.0, "primacy": 1.0, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"E0540S_EUR_NL": 242.0, "components": ["P0733S vial 0.05ml 50000units/ml", "P0720S vial 0.04ml 50000units/ml", "GlycoBuffer 2 1ml 10X", "NP-40 1ml 10%", "Glycoprotein Denaturing Buffer 1ml 10X"], "format": "bundle/1 set"}, "used_in": ["ch07"], "notes": "NEB E0540S欧洲分销价€242；产品组成验证：双酶（O-糖苷酶+神经氨酸酶）+三种缓冲液组件；注：分销商价格非NEB官网直销价，仅作参照"}
{"id": "src_415", "tier": 1, "score": 8.5, "type": "product_catalog", "title": "Genovis SmartEnzymes Store — Complete Product Lineup with Official Pricing 2026", "authors": ["Genovis AB"], "year": 2026, "venue": "Genovis AB Official Store", "url": "https://www.genovis.com/store/", "accessed_at": "2026-04-21", "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"OglyZOR_Lyophilized_2000u_EUR": 1079, "OmniGLYZOR_Kit_EUR_range": "1524-2739", "OpeRATOR_Lyophilized_2000u_EUR": 1251, "ImpaRATOR_Lyophilized_EUR": 1251, "SialEXO_Lyophilized_2000u_EUR": 781, "GlycINATOR_Lyophilized_EUR": 658, "IgGZERO_Lyophilized_EUR": "348-1302"}, "used_in": ["ch07"], "notes": "OglyZOR（O-糖苷酶，SpGH101来源）€1,079/2000u；OmniGLYZOR（N+O糖全套工作流套装）€1,524-2,739；SialEXO€781；为竞品工作流套装定价的核心参照数据"}
{"id": "src_416", "tier": 2, "score": 7.0, "type": "technical_brochure", "title": "GenScript BacPower™ E. coli Protein Expression Service — Technical Handbook & Service Brochure", "authors": ["GenScript Corporation"], "year": 2022, "venue": "GenScript Official Technical Documentation", "url": "https://www.genscript.com/gsfiles/techfiles/B_Recombinant_Protein_Service_Handbook.pdf", "accessed_at": "2026-04-21", "authority": 1.5, "recency": 1.5, "primacy": 1.5, "verifiability": 1.5, "coi": 0.0, "conflict_of_interest": "GenScript官方材料，商业利益相关，降权使用", "blacklist_checked": true, "retraction_checked": false, "key_data": {"max_ecoli_fermentation_volume_L": 2000, "protein_purity_pct": ">=98%", "ecoli_success_rate_pct": 98, "batch_history": ">50000", "BacPower_delivery_weeks": "4", "BacPower_min_delivery_mg": 3, "inclusion_body_platform": "FoldArt™", "cell_pellet_max_tons": 3}, "used_in": ["ch08", "ch12"], "notes": "金斯瑞 BacPower™ 技术手册，明确支持 E. coli 1L-2000L 发酵规模，最大 3 吨湿菌体，克级蛋白纯度 ≥98%；具备包涵体复性平台 FoldArt™；是第 8 章 CDMO 能力矩阵金斯瑞一行的核心来源。"}
{"id": "src_417", "tier": 2, "score": 8.0, "type": "corporate_announcement", "title": "合成生物学业务被市场看好 金斯瑞旗下百斯杰获2.5亿元融资", "authors": ["金斯瑞生物科技股份有限公司（HK:1548）"], "year": 2023, "venue": "金斯瑞官方新闻稿", "url": "https://www.genscript.com.cn/genscript-subsidiary-bestzyme-secured-rmb-250-million-financing-for-synthetic-biology-business.html", "accessed_at": "2026-04-21", "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": "上市公司官方公告，存在正面表述偏向，但融资数据为法律意义上的公开披露", "blacklist_checked": true, "retraction_checked": false, "key_data": {"financing_round": "A轮", "financing_amount_rmb": 250000000, "lead_investor": "高瓴资本", "post_money_valuation_rmb": 2400000000, "genscript_stake_pct": 82.59, "industrial_enzyme_products": 20, "awards": ["全国酶制剂行业十强", "国家级专精特新小巨人", "省高成长中小企业"], "founded": 2013}, "used_in": ["ch08", "ch12"], "notes": "百斯杰 2023 A 轮融资 2.5 亿 RMB，高瓴领投 1 亿，投后估值约 24 亿 RMB；金斯瑞间接持股 82.5917%；百斯杰已开发 20+ 自主知识产权工业酶，全国酶制剂十强企业。"}
{"id": "src_418", "tier": 1, "score": 9.0, "type": "semi_annual_report", "title": "南京诺唯赞生物科技股份有限公司 2025 年半年度报告", "authors": ["南京诺唯赞生物科技股份有限公司"], "year": 2025, "venue": "上交所科创板（688105.SH）", "url": "https://qxb-pdf-osscache.qixin.com/AnBaseinfo/14557464fc8814f446ef6129d742addc.pdf", "accessed_at": "2026-04-21", "authority": 3.0, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 0.5, "conflict_of_interest": "上市公司半年报，存在选择性正面表述可能，但财务数据受证监会监管", "blacklist_checked": true, "retraction_checked": false, "key_data": {"GMP_fermentation_scale_max_L": 100, "mRNA_enzyme_annual_capacity_kg": 250, "mRNA_enzyme_per_batch_kg": 5, "quality_standards": ["中国药品GMP", "ICH Q7", "ICH Q10", "ISO 9001"], "GMP_lines": 2, "total_assets_2025H1_bn_rmb": 5.24}, "used_in": ["ch08", "ch12"], "notes": "诺唯赞龙潭 GMP 车间已运行 GMP 级 10L-100L 发酵线；年产能满足 250 kg mRNA 生产；质量体系参照 GMP/ICH Q7/Q10/ISO 9001；2025H1 通过 SGS 官方复审，是研究级酶 GMP 生产的重要行业参照。"}
{"id": "src_420", "tier": 3, "score": 6.0, "type": "equity_research", "title": "细胞基因治疗CDMO行业深度报告（含CRB 2020行业调查数据，CDMO选择原因分布）", "authors": ["东吴证券研究所"], "year": 2021, "venue": "中国医药工业信息中心/东吴证券", "url": "http://www.cnpharm.com/upload/resources/file/2021/08/04/96042.pdf", "accessed_at": "2026-04-21", "authority": 1.5, "recency": 1.5, "primacy": 1.0, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": "券商研报，可能存在推荐标的利益；但 CRB 数据为独立第三方调查", "blacklist_checked": true, "retraction_checked": false, "key_data": {"cdmo_selection_reason_gmp_pct": 54, "cdmo_selection_reason_capex_pct": 18, "self_build_reason_minimize_total_cost_pct": 46, "cdmo_cost_pct_of_revenue_note": "15-25% 行业均值，生物药CDMO"}, "used_in": ["ch08", "ch12"], "notes": "CRB 2020 调查：54% 企业选 CDMO 首因是'有限 GMP 能力'；CDMO 代工费占收入 15–25% 为行业均值；仅为估算参考，非 O-糖苷酶研究试剂级具体报价。[待验证：需获取专项报价]"}
{"id": "src_421", "tier": 4, "score": 4.5, "type": "engineering_reference", "title": "生物安全实验室建设每平方米造价参考 + ISO 13485认证费用行业参考（美泰诺格实验室工程公司）", "authors": ["美泰诺格工程咨询"], "year": 2023, "venue": "美泰诺格公司官网", "url": "http://www.mtngjh.com/news/detail/831.html", "accessed_at": "2026-04-21", "authority": 0.5, "recency": 2.0, "primacy": 0.5, "verifiability": 0.5, "coi": 0.0, "conflict_of_interest": "工程商网站，存在报价偏高动机", "blacklist_checked": true, "retraction_checked": false, "key_data": {"biosafety_lab_cost_rmb_per_sqm_avg": 2000, "gmp_cleanroom_cost_rmb_per_sqm_range": "2000-6000", "iso13485_certification_cost_rmb_range": "30-80万（行业参考）", "iso13485_certification_months": "12-18"}, "used_in": ["ch08"], "notes": "Tier 4，仅用于量级参考；GMP 建设成本决策时建议向至少 3 家专业工程公司获取正式报价校正。"}
{"id": "src_422", "tier": 2, "score": 7.5, "type": "institutional_recruitment", "title": "中科院上海有机所高级人才招聘启事（糖化学生物学）+ 中科院过程工程研究所糖生物工程课题组招聘", "authors": ["中国科学院上海有机化学研究所", "中国科学院过程工程研究所"], "year": 2023, "venue": "中科院官网", "url": "https://sioc.cas.cn/team/zp/zxzp/202307/t20230719_6812596.html", "accessed_at": "2026-04-21", "authority": 2.5, "recency": 1.5, "primacy": 2.0, "verifiability": 1.5, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"sioc_recruitment_direction": "糖化学生物学方向研究员/课题组长", "ipe_recruitment_direction": "酶催化合成实验员/工程师（人才派遣）", "ipe_url": "https://ipe.cas.cn/rcdw/rczp/202409/t20240902_7331556.html"}, "used_in": ["ch08", "ch12"], "notes": "SIOC 和过程工程所的糖生物学/糖化学人才招聘公告证明该方向人才主要向学术机构聚集；企业需提供 2–3 倍薪酬溢价才能竞争到此类人才。"}
{"id": "src_423", "tier": 2, "score": 7.5, "type": "institutional_recruitment", "title": "中国科学院天津工业生物技术研究所2026年招聘启事（糖生物学/酶工程方向博士后）", "authors": ["中国科学院天津工业生物技术研究所"], "year": 2026, "venue": "优秀人才网（中科院天津工业所授权发布）", "url": "https://www.youxiuhr.com/index.php?c=ads&id=222", "accessed_at": "2026-04-21", "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 1.5, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"research_directions": ["糖生物学", "酶工程", "多酶级联反应"], "special_postdoc_annual_salary_rmb": 340000, "tianjin_annual_subsidy_rmb": 150000, "tianjin_one_time_subsidy_rmb": 50000, "special_postdoc_funding_rmb": "100-150万"}, "used_in": ["ch08", "ch12"], "notes": "天津工业所朱之光课题组招募糖生物学/酶工程博士后（2026年）；特别博士后年薪约 34 万税前（含公积金）+ 天津生活补贴 15 万/年；企业糖生物学 PI 需提供 2–3 倍溢价（80–120 万+股权）才具竞争力。"}
{"id": "src_424", "tier": 4, "score": 4.5, "type": "job_posting_aggregate", "title": "生物医药行业薪酬参考数据（BOSS直聘/领英公开招聘 2024–2026）", "authors": ["BOSS直聘", "LinkedIn"], "year": 2025, "venue": "公开招聘平台", "url": "https://www.zhipin.com", "accessed_at": "2026-04-21", "authority": 0.5, "recency": 2.0, "primacy": 0.5, "verifiability": 0.5, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"fermentation_engineer_3_5yr_rmb_range": "25-45万/年", "qa_manager_5_8yr_rmb_range": "30-50万/年", "pi_biotech_startup_rmb_range": "50-120万/年", "overseas_pi_with_relocation_rmb_range": "80-120万+安家费20-30万"}, "used_in": ["ch08", "ch12"], "notes": "Tier 4，基于公开招聘页面薪酬区间汇总，仅作首年团队薪酬总包量级估算；PI 薪酬区间弹性大，实际取决于候选人背景和谈判。"}
{"id": "src_425", "tier": 2, "score": 6.5, "type": "review", "title": "Challenges Associated With the Formation of Recombinant Protein Inclusion Bodies in Escherichia coli and Strategies to Address Them for Industrial Applications", "authors": ["Garg R", "et al."], "year": 2021, "venue": "Frontiers in Microbiology (PMC7902521)", "doi": null, "pmcid": "PMC7902521", "url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC7902521/", "accessed_at": "2026-04-21", "authority": 2.0, "recency": 1.5, "primacy": 1.5, "verifiability": 1.5, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "key_data": {"inclusion_body_rate_above_80kDa": "50-70%（未优化条件下，综合多篇综述估算）", "strategies": ["低温诱导", "MBP/SUMO融合", "SHuffle菌株", "包涵体复性"]}, "used_in": ["ch09", "ch12"], "notes": "E.coli过表达>80 kDa蛋白时包涵体发生率综合综述；是ch09 T1技术风险评分（概率4/5）的文献支撑；2021年综述，时效性可接受"}
{"id": "src_426", "tier": 1, "score": 8.0, "type": "journal", "title": "Enhanced Recombinant Protein Production of Soluble, Highly Active and Immobilizable PNGase F", "authors": ["Szarka A", "et al."], "year": 2022, "venue": "Biomolecules (PMC9259526)", "doi": null, "pmcid": "PMC9259526", "url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC9259526/", "accessed_at": "2026-04-21", "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "key_data": {"protein": "PNGase F (37 kDa)", "activity_storage": "stable 24 months at -20°C in 50% glycerol", "clone_to_activity_note": "类比GH101推算8-14周工艺周期"}, "used_in": ["ch09"], "notes": "PNGase F重组表达优化案例；作为GH101开发时间表和活性验证周期的类比参照；2022年Biomolecules（MDPI，IF约4.0）。注：PNGase F（37 kDa）远小于EngEF（108 kDa），类比须谨慎。"}
{"id": "src_427", "tier": 1, "score": 9.5, "type": "regulatory", "title": "Provisions for In-Vitro Diagnostic Reagent Registration and Filing (NMPA Order No.48, 2021) — English version", "authors": ["NMPA China"], "year": 2021, "venue": "NMPA Official English Portal", "doi": null, "url": "https://english.nmpa.gov.cn/2024-06/05/c_1049322.htm", "accessed_at": "2026-04-21", "authority": 3.0, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"class_I_requirement": "filing only, no technical review", "class_II_timeline_months": "12-22", "RUO_exemption": "仅供研究不用于诊断产品豁免注册，须标注RUO字样", "effective_date": "2021-10-01"}, "used_in": ["ch09"], "notes": "NMPA 2021年第48号令最新有效文本；IVD Class I仅需备案；RUO产品豁免注册是O-糖苷酶科研试剂18个月路径的关键合规依据；英文版本核查补充：中文版见国家药监局官网，英文版2024-06-05发布"}
{"id": "src_428", "tier": 2, "score": 6.5, "type": "industry_article", "title": "CDMO selection: Startups need patience, humility, and good faith", "authors": ["Abbott J", "BioProcess International"], "year": 2024, "venue": "BioProcess International", "doi": null, "url": "https://www.bioprocessintl.com/deal-making/cdmo-selection-startups-need-patience-humility-and-good-faith", "accessed_at": "2026-04-21", "authority": 1.5, "recency": 2.0, "primacy": 1.5, "verifiability": 1.5, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"key_finding": "CDMO合作成功依赖信任和持续沟通；放大失败和批间一致性是常见问题；建议初创企业与有经验CDMO合作即使成本更高", "conference": "BPI Europe 2024"}, "used_in": ["ch09"], "notes": "BPI行业媒体文章，BPI Europe 2024会议报道；支持双CDMO布局策略和T3批间一致性风险评分逻辑"}
{"id": "src_429", "tier": 3, "score": 5.5, "type": "industry_article", "title": "5 Reasons Why Biotech Startups Fail: How to Avoid Them", "authors": ["Labiotech editorial team"], "year": 2023, "venue": "Labiotech.eu", "doi": null, "url": "https://www.labiotech.eu/members/biotech-startups-failure-advice/", "accessed_at": "2026-04-21", "authority": 1.0, "recency": 2.0, "primacy": 1.0, "verifiability": 1.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"key_finding": "失败越晚越贵，应在早期设置Go/No-Go决策点；36%生物技术公司在初期阶段退出", "quote": "Failure is just more painful and expensive the later it happens"}, "used_in": ["ch09"], "notes": "Tier3行业媒体；仅用于支持Kill Criteria设置的管理逻辑合理性，不作为核心技术结论唯一支撑"}
{"id": "src_430", "tier": 1, "score": 9.2, "type": "journal", "title": "Structural basis of mammalian mucin processing by the human gut O-glycopeptidase OgpA from Akkermansia muciniphila", "authors": ["Trastoy B", "Naegeli A", "Anso I", "Sjogren J", "Guerin ME"], "year": 2020, "venue": "Nature Communications", "doi": "10.1038/s41467-020-18696-y", "pmid": "32973204", "pmcid": "PMC7518263", "url": "https://www.nature.com/articles/s41467-020-18696-y", "accessed_at": "2026-04-21", "authority": 3.0, "recency": 1.6, "primacy": 2.0, "verifiability": 2.0, "coi": 0.5, "conflict_of_interest": "Naegeli和Sjogren为Genovis AB员工，Naegeli持有公司股份；但结构数据独立可验证", "blacklist_checked": true, "retraction_checked": true, "key_data": {"enzyme": "OgpA (OpeRATOR)", "source_organism": "Akkermansia muciniphila", "molecular_weight_kDa": 42, "expression_host": "E. coli His-tagged", "mechanism": "水解O-糖基化Ser/Thr残基N端肽键", "substrate_pref": "asialylated Core 1 O-glycans", "commercial_name": "OpeRATOR® (Genovis AB)", "citations": 102, "pdb_id": "6Z2P"}, "used_in": ["ch10"], "notes": "OpeRATOR结构解析奠基论文；2020年Nat Commun IF~17，102次引用；Trastoy/Guerin为学术作者，结构数据独立；利益冲突已声明，评分降0.5分；是OpeRATOR商品化的科学基石"}
{"id": "src_431", "tier": 2, "score": 7.0, "type": "commercial", "title": "Genovis OpeRATOR Lyophilized / ImpaRATOR Lyophilized — Product Pages and Annual Report 2024", "authors": ["Genovis AB"], "year": 2024, "venue": "Genovis official website + MFN Annual Report 2024", "url": "https://www.genovis.com/smartenzymes/glycan-profiling/operator/", "accessed_at": "2026-04-21", "authority": 2.0, "recency": 2.0, "primacy": 1.5, "verifiability": 1.5, "coi": 0.0, "conflict_of_interest": "Genovis是这些产品的销售方，定价和规格数据存在商业利益", "blacklist_checked": true, "retraction_checked": false, "key_data": {"OpeRATOR_price_EUR": 1251, "OpeRATOR_size": "2000 units (2 mg protein)", "OpeRATOR_MW_kDa": 42, "OpeRATOR_substrate": "core 1 O-glycans, requires SialEXO for sialylated substrates", "SialEXO_price_EUR": 781, "ImpaRATOR_price_EUR": 1251, "ImpaRATOR_MW_kDa": 97, "ImpaRATOR_source": "Pseudomonas aeruginosa expressed in E. coli", "ImpaRATOR_substrate": "broad including sialylated Core 1 and Core 2, Tn antigen", "ImpaRATOR_adjacent_limitation": "limited activity at sites with two adjacent glycosylated Ser/Thr", "AR2024_new_products": 6, "AR2024_new_patent_filings": "several new enzymes"}, "used_in": ["ch10"], "notes": "Genovis官网产品页（2025年检索）及2024年年报；产品参数直接来自商业页面，是定价和规格的第一手数据；需注意商业利益因素，但技术规格无动机虚报"}
{"id": "src_432", "tier": 1, "score": 9.0, "type": "journal", "title": "Glycoproteomic landscape and structural dynamics of TIM family immune checkpoints enabled by mucinase SmE", "authors": ["Chongsaritsinsuk J", "Steigmeyer AD", "Mahoney KE", "Rosenfeld MA", "Lucas TM", "Smith CM", "Malaker SA", "et al."], "year": 2023, "venue": "Nature Communications", "doi": "10.1038/s41467-023-41756-y", "pmid": "37794035", "url": "https://www.nature.com/articles/s41467-023-41756-y", "accessed_at": "2026-04-21", "authority": 3.0, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 0.5, "conflict_of_interest": "Malaker/Shon/Bertozzi是Stanford mucinase专利共同发明人；但数据独立可验证", "blacklist_checked": true, "retraction_checked": true, "key_data": {"SmE_source": "Serratia marcescens (M60-like metallopeptidase)", "SmE_MW_kDa": 94, "SmE_advantage": "cleaves at complex glycans and adjacent O-glycosites, no sialidase required", "OgpA_glycosites_identified": 113, "SmE_glycosites_vs_OgpA": ">2x more than OgpA", "IMPa_adjacent_limitation": "does not cleave between two adjacent glycosylated residues (confirmed)", "benchmark_proteins": "TIM-1, TIM-3, TIM-4 immune checkpoints", "TIM3_vs_TIM1_TIM4_glycosites": "TIM-3 has markedly fewer O-glycosites"}, "used_in": ["ch10"], "notes": "SmE定义性论文；首次完成TIM家族完整O-糖组学图谱；提供OgpA/IMPa/SmE三款酶的直接基准测试数据；2023年Nat Commun，是迄今O-糖肽酶领域覆盖面最广的比较研究"}
{"id": "src_433", "tier": 1, "score": 8.8, "type": "journal", "title": "A Broad-Specificity O-Glycoprotease That Enables Improved Analysis of Glycoproteins and Glycopeptides Containing Intact Complex O-Glycans", "authors": ["Vainauskas S", "Duke RM", "McFarland M", "McClung C", "Taron CH"], "year": 2022, "venue": "Analytical Chemistry", "doi": "10.1021/acs.analchem.1c04055", "pmid": "34962767", "url": "https://pubs.acs.org/doi/10.1021/acs.analchem.1c04055", "accessed_at": "2026-04-21", "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 0.5, "conflict_of_interest": "Vainauskas和Taron均来自NEB（New England Biolabs），具有IMPa商业利益；数据已独立验证", "blacklist_checked": true, "retraction_checked": true, "key_data": {"enzyme": "IMPa", "source_organism": "Pseudomonas aeruginosa (CpaA homolog)", "sialic_acid_tolerance": "yes - cleaves sialylated Core 1, sialylated Core 2, Tn antigen", "motif_preference": "Pro-Ser/Thr motif (proline at P1 position)", "no_adjacent_glycosite_cleavage": true, "application": "one-step O-glycoproteomic workflow for G-CSF glycoprofiling", "commercial_product": "ImpaRATOR (Genovis, also NEB-distributed)"}, "used_in": ["ch10"], "notes": "IMPa定义性论文；NEB内部研究但发表于ACS Analytical Chemistry（IF~8.0）；奠定ImpaRATOR商品化基础；NEB利益冲突已声明，但方法学独立性可接受；sialic acid tolerance是O-糖肽酶领域里程碑突破"}
{"id": "src_434", "tier": 1, "score": 8.5, "type": "journal", "title": "Structural evidence for a proline-specific glycopeptide recognition domain in an O-glycopeptidase", "authors": ["Noach I", "Boraston AB"], "year": 2021, "venue": "Glycobiology", "doi": "10.1093/glycob/cwaa102", "pmid": "33030205", "pmcid": "PMC8091461", "url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC8091461/", "accessed_at": "2026-04-21", "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "key_data": {"enzyme": "IMPa", "structure": "crystal structure with O-glycopeptide complex (PDB: 5KDX)", "IMPa_N2_domain": "proline recognition domain with 4-aromatic bowl binding Pro-Ser/Thr-O-glycan motif", "mechanism": "aromatic bowl accommodates proline; GalNAc wraps around Tyr residue via CH-π interaction", "comparison_to_OgpA": "Tyr116 of OgpA binds Gal rather than Sia, explaining sialic acid sensitivity contrast"}, "used_in": ["ch10"], "notes": "Noach/Boraston（UBC）独立学术研究，无利益冲突；揭示IMPa唾液酸耐受的结构基础，为工程改造提供靶点；2021年Glycobiology（IF~4.7）"}
{"id": "src_435", "tier": 1, "score": 9.0, "type": "journal", "title": "Targeted glycan degradation potentiates the anticancer immune response in vivo", "authors": ["Gray MA", "Stanczak MA", "Mantuano NR", "Xiao H", "Pijnenborg JFA", "Malaker SA", "Bertozzi CR", "et al."], "year": 2020, "venue": "Nature Chemical Biology", "doi": "10.1038/s41589-020-0622-x", "pmid": "32989299", "url": "https://www.nature.com/articles/s41589-020-0622-x", "accessed_at": "2026-04-21", "authority": 3.0, "recency": 1.5, "primacy": 2.0, "verifiability": 2.0, "coi": 0.5, "conflict_of_interest": "Bertozzi是Palleon创始人，有商业利益；但学术论文数据独立", "blacklist_checked": true, "retraction_checked": true, "key_data": {"strategy": "antibody-enzyme conjugate with sialidase for glycocalyx editing", "result": "potentiated anti-tumor immunity in vivo", "journal_IF_approx": 14, "significance": "first in vivo proof of targeted glycocalyx degradation for cancer immunotherapy"}, "used_in": ["ch10"], "notes": "Gray/Bertozzi 2020 Nat Chem Biol；确立靶向糖萼降解促肿瘤免疫的体内概念验证，是eStcE治疗化研究的直接前驱；Palleon E-602的学术基础之一"}
{"id": "src_437", "tier": 1, "score": 8.0, "type": "patent", "title": "Stanford University Technology - Cell-type specific enzymatic degradation of pathological mucins (eStcE fusion construct, WO2023212733)", "authors": ["Stanford University - Office of Technology Licensing"], "year": 2023, "venue": "USPTO / WIPO", "url": "https://techfinder.stanford.edu/technology/cell-type-specific-enzymatic-degradation-pathological-mucins", "accessed_at": "2026-04-21", "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 1.5, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"patent_WO": "WO2023212733", "US_published_application": "US20250276081", "core_claim_scope": "fusion constructs of engineered mucinase + cancer antigen-targeting nanobody/antibody", "inventors": "Pedram K, Shon DJ, Tender GS, Bertozzi CR et al.", "Stanford_docket": "Stanford Technology License"}, "used_in": ["ch10"], "notes": "Stanford官方技术转让页面；WO2023212733专利核心权利要求针对'融合构型'（eStcE+靶向纳米抗体），eStcE酶本体的单独使用和非HER2靶向构型是否被覆盖需进一步检索full claims；是判断IP空白空间的关键文件"}
{"id": "src_438", "tier": 1, "score": 9.0, "type": "clinical_trial", "title": "Palleon Pharmaceuticals E-602 (HLX79) Phase 1/2 GLIMMER-01 + Phase 2 Glomerulonephritis Trial", "authors": ["Palleon Pharmaceuticals"], "year": 2022, "venue": "ClinicalTrials.gov + Palleon Press Releases", "url": "https://palleonpharma.com/press-releases/palleon-pharmaceuticals-announces-first-patient-dosed-in-phase-1-2-study-of-lead-glyco-immunology-drug-candidate-for-oncology/", "accessed_at": "2026-04-21", "authority": 3.0, "recency": 2.0, "primacy": 2.0, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": "Palleon是商业公司，但IND/临床进展是公开官方事实", "blacklist_checked": true, "retraction_checked": false, "key_data": {"E602_IND_clearance": "2022-01-20 (FDA IND cleared)", "GLIMMER01_first_patient": "2022-03-08", "NCT": "NCT05259696", "GLIMMER01_design": "Phase 1/2, solid tumors, sialidase-Fc E-602 + cemiplimab", "Phase2_glomerulonephritis_first_patient": "2025-08-07", "Phase2_compound": "E-602 (HLX79) + rituximab biosimilar", "China_NMPA_IND": "cleared for Phase 2", "Palleon_co_founder": "Carolyn Bertozzi (2015)", "series_B": "100 million USD", "AACR2026_abstract": "HLX316/E-688 anti-B7-H3 sialidase"}, "used_in": ["ch10"], "notes": "Palleon E-602是Bertozzi实验室糖萼编辑疗法进入临床的第一个分子（唾液酸酶，非mucinase），但已证明微生物来源工程糖苷酶的人体注射安全性概念；对eStcE治疗化路径有重要参考意义；IND窗口预估依据Palleon从基础研究(2015)到Phase1(2022)约7年的时间线"}
{"id": "src_439", "tier": 1, "score": 8.5, "type": "journal", "title": "The protease cathepsin K can debulk the cancer glycocalyx", "authors": ["Bertozzi CR group", "et al."], "year": 2026, "venue": "Journal of Biological Chemistry (PubMed ID: 41581870)", "pmid": "41581870", "url": "https://pubmed.ncbi.nlm.nih.gov/41581870/", "accessed_at": "2026-04-21", "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 0.5, "conflict_of_interest": "Bertozzi多家公司创始人；但学术独立数据", "blacklist_checked": true, "retraction_checked": false, "key_data": {"finding": "CTSK (cathepsin K, human enzyme) uniquely degrades cell-surface mucins, proteoglycans, polysialylated glycoproteins", "comparison": "screened all 15 human cathepsins", "patent": "Stanford STAN-2144WO filed", "significance": "humanized alternative to bacterial mucinase eStcE, addresses immunogenicity concern"}, "used_in": ["ch10"], "notes": "2026年JBC；提示治疗化mucinase正向人源化方向演进，缓解细菌来源免疫原性担忧；与eStcE构成技术竞争路径；是判断'治疗化赛道仍早期'的最新数据点"}
{"id": "src_441", "tier": 2, "score": 7.5, "type": "review", "title": "Carbohydrate-active enzyme (CAZyme) discovery and engineering via (Ultra)high-throughput screening", "authors": ["Wardman JF", "Withers SG"], "year": 2024, "venue": "RSC Chemical Biology", "doi": "10.1039/D4CB00024B", "url": "https://pubs.rsc.org/en/content/articlehtml/2024/cb/d4cb00024b", "accessed_at": "2026-04-21", "authority": 2.0, "recency": 2.0, "primacy": 1.5, "verifiability": 1.5, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "key_data": {"MELiORA_summary": "MELiORA listed as most promising uHTS platform for CAZyme engineering", "application_range": "O-glycopeptidases, glycosyltransferases, glycoside hydrolases"}, "used_in": ["ch10", "ch12"], "notes": "Wardman/Withers 2024 RSC Chem Biol综述；MELiORA平台扩展应用综述；是MELiORA成为CAZyme工程标准方法的佐证文献"}
{"id": "src_442", "tier": 1, "score": 8.5, "type": "journal", "title": "Reshaping of a Glycoside Hydrolase Active Site through Expression-Compensated Droplet-Based Microfluidic Screening Provides Useful Tools for Glycomics", "authors": ["Wardman JF", "Withers SG", "et al."], "year": 2025, "venue": "ACS Central Science", "doi": "10.1021/acscentsci.5c01227", "pmid": "41142332", "url": "https://pubs.acs.org/doi/full/10.1021/acscentsci.5c01227", "accessed_at": "2026-04-21", "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"target": "SpGH101 Q868G variant directed evolution", "method": "expression-compensated droplet-based microfluidic FACS", "substrate": "sialyl T-antigen (Neu5Ac-Gal-GalNAc-Ser/Thr)", "result": "improved variants with enhanced sialyl T-antigen cleavage activity beyond Q868G"}, "used_in": ["ch10", "ch12"], "notes": "Wardman/Withers 2025 ACS Central Science；对Q868G骨架进行液滴微流控定向进化的最新成果；证明单点突变可进一步工程化；是双路线IP布局中路线一的关键科学支撑；2025年最新发表"}
{"id": "src_443", "tier": 2, "score": 7.5, "type": "review", "title": "AlphaFold3 — Accurate Structure Prediction of Biomolecular Interactions (Nature 2024) and AI-Driven De Novo Protein Design in Enzyme Engineering", "authors": ["Abramson J", "et al.", "DeepMind"], "year": 2024, "venue": "Nature (AlphaFold3) + Biology MDPI review", "doi": "10.1038/s41586-024-07487-w", "url": "https://www.nature.com/articles/s41586-024-07487-w", "accessed_at": "2026-04-21", "authority": 3.0, "recency": 2.0, "primacy": 1.5, "verifiability": 1.5, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"AlphaFold3_release": "2024-05-08", "RoseTTAFold_All_Atom_release": "2024-03-07", "capability": "predict biomolecular complexes including proteins, small molecules, modified residues", "application_to_CAZymes": "multiple labs use AF3 for carbohydrate-active enzyme structure analysis and virtual mutant screening"}, "used_in": ["ch10"], "notes": "AlphaFold3 Nature 2024原文；结合MDPIreview（Biology 2025，14:1268）对AI蛋白设计在酶工程中应用的综述；AF3是工程酶赛道加速的关键工具；降低理性设计成本数量级"}
{"id": "src_445", "tier": 2, "score": 7.0, "type": "legal_reference", "title": "PCT Filing Costs and Fees: Complete Breakdown — Teak IP Services / USPTO PCT Fees in USD", "authors": ["Teak IP Services", "USPTO"], "year": 2024, "venue": "USPTO Official + Teak IP Services", "url": "https://www.uspto.gov/patents/basics/international-protection/patent-cooperation-treaty/pct-fees-us-dollars", "accessed_at": "2026-04-21", "authority": 2.0, "recency": 2.0, "primacy": 1.5, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": "Teak IP为专利服务公司，有推荐服务动机；但USPTO官方费率无利益冲突", "blacklist_checked": true, "retraction_checked": false, "key_data": {"PCT_international_filing_fee_USD": 1667, "PCT_search_fee_approx_USD": "1000-2000 (depending on ISA)", "PCT_international_phase_total_USD": "5000-10000", "national_phase_total_per_country_USD": "3000-8000", "complete_3country_protection_USD": "15000-50000 per patent", "RMB_estimate_2PCT_full_global": "100-150万RMB (含律师费+国家阶段)"}, "used_in": ["ch10", "ch12"], "notes": "PCT费率参考；USPTO官方费率为基准，Teak IP提供完整成本分解；用于第12章预算支撑；利益冲突注意但数量级可信"}
{"id": "src_446", "tier": 1, "score": 9.0, "type": "journal", "title": "Prevalence and trend of biopsy-proven IgA nephropathy in China: a systematic review", "authors": ["Zhang et al."], "year": 2025, "venue": "Journal of Nephrology (Springer)", "doi": "10.1007/s40620-025-02265-3", "pmid": "40138167", "url": "https://pubmed.ncbi.nlm.nih.gov/40138167/", "accessed_at": "2026-04-21", "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"total_patients": 143176, "IgAN_proportion": "39.73%", "IgAN_cases": 56886, "provinces_covered": 34, "male_female_ratio": "58.67% / 41.33%", "weighted_avg_pre2010": "40.45%", "weighted_avg_post2010": "37.97%"}, "used_in": ["ch11"], "notes": "2025年发表的中国IgAN最全面系统综述，覆盖34个省市143,176例原发性肾小球疾病活检数据；IgAN占39.73%，是中国最常见原发性肾小球疾病。Italian Society of Nephrology旗下期刊。Tier 1学术文献。"}
{"id": "src_447", "tier": 2, "score": 7.5, "type": "market_report", "title": "IgA Nephropathy Epidemiology and Market Forecast 2034 — China & 7MM", "authors": ["DelveInsight Business Research"], "year": 2024, "venue": "DelveInsight", "url": "https://www.delveinsight.com/report-store/iga-nephropathy-epidemiology-forecast", "accessed_at": "2026-04-21", "authority": 2.0, "recency": 2.0, "primacy": 1.0, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"global_7MM_China_prevalent_2022": 1899000, "japan_diagnosed_2024": 175000, "US_diagnosed_2024": 133000, "US_IgAN_market_2024_USD_M": 455, "EU4_UK_market_2024_USD_M": 150, "Japan_market_2024_USD_M": 125}, "used_in": ["ch11"], "notes": "DelveInsight为Tier 2医疗市场报告，2024年数据；7MM+中国IgAN患者约190万例（2022年）；中国实际患者规模需考虑活检率不足因素，为中国IgAN市场规模的主要公开引用来源"}
{"id": "src_448", "tier": 1, "score": 9.0, "type": "regulatory", "title": "China Government Procurement Domestic Product Standards — Ministry of Finance/MIIT Notice (effective 2026-01-01)", "authors": ["Ministry of Finance of China", "Ministry of Industry and Information Technology"], "year": 2025, "venue": "www.gov.cn (Official China Government Portal)", "url": "https://english.www.gov.cn/news/202512/19/content_WS69454fa9c6d00ca5f9a0834b.html", "accessed_at": "2026-04-21", "authority": 3.0, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"effective_date": "2026-01-01", "price_preference_pct": 20, "eligible_entity": "both domestic and foreign-invested enterprises meeting domestic product criteria", "definition": "manufactured or substantially transformed in China", "medical_device_criterion": "NMPA domestic device registration certificate = domestic product"}, "used_in": ["ch11"], "notes": "2025年12月19日中国财政部+工信部联合发布，2026年1月1日起国产品在政府采购中享有20%评价价格优惠；医疗器械以NMPA国产注册证为'国产'认定依据；是支持供应链轴政策东风的Tier 1监管文件"}
{"id": "src_449", "tier": 1, "score": 9.5, "type": "clinical_guideline", "title": "KDIGO 2025 Clinical Practice Guideline for the Management of IgA Nephropathy and IgA Vasculitis", "authors": ["Kidney Disease: Improving Global Outcomes (KDIGO) IgAN Work Group"], "year": 2025, "venue": "Kidney International (Supplement)", "doi": "10.1016/j.kint.2025.xx", "url": "https://kdigo.org/wp-content/uploads/2024/08/KDIGO-2025-IgAN-IgAV-Guideline.pdf", "accessed_at": "2026-04-21", "authority": 3.0, "recency": 2.0, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"diagnosis_standard": "kidney biopsy gold standard; NO validated serum/urine biomarker for diagnosis", "east_asian_highest_prevalence": true, "incidence_global": "2.5 per 100,000 per year", "progression_rate": "30-40% ESRD within 20-30 years"}, "used_in": ["ch11"], "notes": "KDIGO 2025 IgAN/IgAV临床实践指南；明确'目前无经过验证的血清或尿液诊断标志物'；肾活检仍是诊断金标准；东亚裔最高发病率；全球最权威IgAN管理指南，Tier 1监管/临床文献"}
{"id": "src_450", "tier": 2, "score": 8.0, "type": "clinical_lab", "title": "Gd-IgA1 Biomarker Screening Service — Cincinnati Children's Hospital Nephrology Clinical Laboratory", "authors": ["Cincinnati Children's Hospital Medical Center"], "year": 2025, "venue": "Cincinnati Children's Hospital (CLIA/CAP certified clinical laboratory)", "url": "https://www.cincinnatichildrens.org/-/media/Cincinnati-Childrens/Home/clinical-labs/nephrology/Gd-IgA1-Biomarker-Screening.pdf", "accessed_at": "2026-04-21", "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 1.5, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": false, "key_data": {"diagnostic_cutoff_ug_mL": 83.2, "AUC": 0.95, "sensitivity_pct": 82.9, "specificity_pct": 91.6, "diagnostic_efficiency_pct": 89.8, "assay_type": "High-sensitivity ELISA, CAP/CLIA/GCLP validated", "method": "IgA-capture reagent + lectin-based galactose-deficient O-glycan detection"}, "used_in": ["ch11"], "notes": "Cincinnati Children's于2025年推出临床Gd-IgA1检测服务，AUC=0.950；CLIA/CAP/GCLP认证；是当前最高规格临床Gd-IgA1实验室检测的性能标杆；印证高特异性诊断试剂需求已经出现"}
{"id": "src_451", "tier": 1, "score": 8.5, "type": "press_release", "title": "Everest Medicines: NMPA Full Approval of NEFECON for IgA Nephropathy (May 2025)", "authors": ["Everest Medicines (HKEX: 1952.HK)"], "year": 2025, "venue": "ACN Newswire / Everest Medicines IR", "url": "https://www.acnnewswire.com/press-release/english/99552/everest-medicines-announces-nmpa-full-approval-of-nefecon,-broadening-treatment-access-for-iga-nephropathy-patients-in-china", "accessed_at": "2026-04-21", "authority": 2.5, "recency": 2.0, "primacy": 2.0, "verifiability": 1.5, "coi": 0.5, "conflict_of_interest": "Everest Medicines为NEFECON持有者，有正面叙事动机；但NMPA批准为客观监管事实", "blacklist_checked": true, "retraction_checked": false, "key_data": {"NMPA_approval": "Full approval (sNDA) May 2025", "indication": "primary IgAN, eGFR≥30 mL/min regardless of proteinuria", "China_annual_new_cases": ">100,000", "NRDL_provinces": 31, "first_etiological_treatment_China": true}, "used_in": ["ch11"], "notes": "NEFECON获NMPA全批准，扩大至不限蛋白尿水平的成人IgAN患者；中国每年新确诊IgAN逾10万例；已纳入31省市NRDL；印证IgAN诊疗市场快速标准化，对诊断试剂需求产生正向拉动"}
{"id": "src_452", "tier": 1, "score": 9.2, "type": "journal", "title": "Analysis of O-glycoforms of the IgA1 hinge region by sequential deglycosylation", "authors": ["Renfrow MB", "Novak J", "et al."], "year": 2020, "venue": "Scientific Reports (Nature Publishing Group)", "doi": "10.1038/s41598-020-57510-z", "pmcid": "PMC6971281", "url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC6971281/", "accessed_at": "2026-04-21", "authority": 2.5, "recency": 1.5, "primacy": 2.0, "verifiability": 2.0, "coi": 1.0, "conflict_of_interest": null, "blacklist_checked": true, "retraction_checked": true, "key_data": {"workflow": "neuraminidase + O-glycanase sequential deglycosylation for IgA1 HR O-glycoform analysis", "EngEF_vs_SpGH101": "EngEF superior: 0.42% disaccharides remaining vs 63.94% for SpGH101 after O-glycanase treatment", "application": "LC-MS based Gd-IgA1 site-specific quantification", "IgA1_HR_O_glycosites": "up to 6 clustered O-glycans on hinge region"}, "used_in": ["ch11"], "notes": "Renfrow/Novak 2020 Sci Rep；确立顺序脱糖（神经氨酸酶→O-糖苷酶）是IgA1铰链区Gd-O-糖型定量分析的标准工作流；EngEF在此工作流中优于SpGH101（残余二糖0.42% vs 63.94%）；是O-糖苷酶进入IgAN诊断市场的核心技术依据"}
