Academic Journal
Glycosylation-based distinction of endogenous and recombinant complement factor H enables selective quantification in human plasma.
| Τίτλος: | Glycosylation-based distinction of endogenous and recombinant complement factor H enables selective quantification in human plasma. |
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| Συγγραφείς: | Schäfer A; Swiss BioQuant AG, Reinach, Switzerland., Dabrowska-Schlepp P; Eleva GmbH, Freiburg, Germany., Busch A; Eleva GmbH, Freiburg, Germany., Chen JY; Swiss BioQuant AG, Reinach, Switzerland., Wiesener M; Department of Nephrology and Hypertension, University Hospital Erlangen, Erlangen, Germany., Siethoff C; Swiss BioQuant AG, Reinach, Switzerland. |
| Πηγή: | Bioanalysis [Bioanalysis] 2026 Aug-Aug; Vol. 18 (12-13), pp. 1095-1106. Date of Electronic Publication: 2026 Oct 01. |
| Τύπος έκδοσης: | Journal Article |
| Γλώσσα: | English |
| Στοιχεία περιοδικού: | Publisher: Taylor & Francis Country of Publication: England NLM ID: 101512484 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1757-6199 (Electronic) Linking ISSN: 17576180 NLM ISO Abbreviation: Bioanalysis Subsets: MEDLINE |
| Imprint Name(s): | Publication: 2024- : [Milton Park, Oxfordshire] : Taylor & Francis Original Publication: London : Future Science, 2009- |
| Ιατρικοί όροι (MeSH): | Complement Factor H*/metabolism , Complement Factor H*/analysis , Complement Factor H*/chemistry, Recombinant Proteins/blood ; Recombinant Proteins/chemistry ; Recombinant Proteins/metabolism ; Chromatography, High Pressure Liquid/methods ; Tandem Mass Spectrometry/methods ; Humans ; Glycosylation |
| Περίληψη: | Background: Therapeutic proteins constitute an important class of biologics, including recombinant coagulation factors, enzyme-replacement therapies and hormones. Generally, a high degree of structural similarity between endogenous and therapeutic proteins is essential. From a pharmacokinetic perspective, however, it is advantageous to selectively quantify the exogenous protein. Research Design: This study presents a strategy for the specific quantification of CPV 104, a recombinant form of complement factor H (FH), in human plasma. Because CPV 104 is identical in amino acid sequence to endogenous FH, differential HPLC-MS/MS analysis was used to identify structural features that discriminate both forms. Results: Differential analysis revealed unique glycosylation patterns on CPV 104, enabling development of a targeted HPLC-MS/MS assay for CPV 104-specific glycopeptides. Assay robustness was further enhanced by incorporating CPV 104 with stable isotope labeled glycan residues as an internal standard. High-throughput analysis was achieved through integration with an affinity enrichment step using a custom FH specific resin, supporting 96 well processing. The method was fully validated and demonstrated excellent specificity, precision, and accuracy. The validated assay was applied to a first in human pharmacokinetics study while simultaneously monitoring endogenous FH. Conclusions: This work outlines a generalizable approach for identifying molecular differences between endogenous and recombinant proteins and leveraging these features to develop high-quality bioanalytical assays. |
| Contributed Indexing: | Keywords: C3 glomerulopathy; HPLC-MS/MS; affinity capture; bioanalysis; glycopeptides; glycosylation; navigo proteins; targeted protein quantification |
| Substance Nomenclature: | 0 (Recombinant Proteins) 80295-65-4 (Complement Factor H) |
| Entry Date(s): | Date Created: 20261001 Date Completed: 20261005 Latest Revision: 20261009 |
| Update Code: | 20261009 |
| PubMed Central ID: | PMC13644591 |
| DOI: | 10.1080/17576180.2026.2740919 |
| PMID: | 42821580 |
| Βάση Δεδομένων: | MEDLINE |
| ISSN: | 1757-6199 |
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| DOI: | 10.1080/17576180.2026.2740919 |