Complete mapping of disulfide linkages for etanercept products by multi-enzyme digestion coupled with LC-MS/MS using multi-fragmentations including CID and ETD

Li Juan Huang, Chia Wang Chiang, Shun Li Chen, Shih Yao Wei, Shu-Hui Chen

Research output: Contribution to journalArticle

Abstract

The disulfide linkages of two etanercept products, Enbrel® (innovator drug) and TuNEX®, were characterized and compared using a multi-fragmentation approach consisting of electron transfer dissociation (ETD) and collision induced dissociation (CID) in combination with multi-enzyme digestion protocols (from Lys-C, trypsin, Glu-C, and PNGase F). Multi-fragmentation approach allowed multi-disulfide linkages contained in a peptide to be un-ambiguously assigned based on the cleavage of both the disulfide and the backbone linkages in a MS 3 schedule. New insights gained using this approach were discussed. A total of 29 disulfides, Cys18-Cys31, Cys32-Cys45, Cys35-Cys53, Cys56-Cys71, Cys74-Cys88, Cys78-Cys-96, Cys98-Cys104, Cys112-Cys121, Cys115-Cys139, Cys-142-Cys157, Cys163-Cys178 in TNFR portion and Cys240-Cys240, Cys246-Cys246, Cys249-Cys249, Cys281-Cys341, Cys387-Cys445 in IgG1 Fc domain, were completely assigned with the demonstration of the same disulfide linkages between the Enbrel® and TuNEX® products. The data showed the higher order structure was preserved throughout the recombinant manufacturing processes and consistent between the two products.

Original languageEnglish
Pages (from-to)531-541
Number of pages11
JournalJournal of Food and Drug Analysis
Volume27
Issue number2
DOIs
Publication statusPublished - 2019 Apr 1

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Chromosome Mapping
sulfides
Disulfides
electron transfer
Digestion
digestion
Electrons
Enzymes
enzymes
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
Trypsin
trypsin
Appointments and Schedules
manufacturing
Immunoglobulin G
Etanercept
tandem mass spectrometry
peptides
drugs
Peptides

All Science Journal Classification (ASJC) codes

  • Food Science
  • Pharmacology

Cite this

@article{cff1c175edcf43429ca13694f77f4707,
title = "Complete mapping of disulfide linkages for etanercept products by multi-enzyme digestion coupled with LC-MS/MS using multi-fragmentations including CID and ETD",
abstract = "The disulfide linkages of two etanercept products, Enbrel{\circledR} (innovator drug) and TuNEX{\circledR}, were characterized and compared using a multi-fragmentation approach consisting of electron transfer dissociation (ETD) and collision induced dissociation (CID) in combination with multi-enzyme digestion protocols (from Lys-C, trypsin, Glu-C, and PNGase F). Multi-fragmentation approach allowed multi-disulfide linkages contained in a peptide to be un-ambiguously assigned based on the cleavage of both the disulfide and the backbone linkages in a MS 3 schedule. New insights gained using this approach were discussed. A total of 29 disulfides, Cys18-Cys31, Cys32-Cys45, Cys35-Cys53, Cys56-Cys71, Cys74-Cys88, Cys78-Cys-96, Cys98-Cys104, Cys112-Cys121, Cys115-Cys139, Cys-142-Cys157, Cys163-Cys178 in TNFR portion and Cys240-Cys240, Cys246-Cys246, Cys249-Cys249, Cys281-Cys341, Cys387-Cys445 in IgG1 Fc domain, were completely assigned with the demonstration of the same disulfide linkages between the Enbrel{\circledR} and TuNEX{\circledR} products. The data showed the higher order structure was preserved throughout the recombinant manufacturing processes and consistent between the two products.",
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Complete mapping of disulfide linkages for etanercept products by multi-enzyme digestion coupled with LC-MS/MS using multi-fragmentations including CID and ETD. / Huang, Li Juan; Chiang, Chia Wang; Chen, Shun Li; Wei, Shih Yao; Chen, Shu-Hui.

In: Journal of Food and Drug Analysis, Vol. 27, No. 2, 01.04.2019, p. 531-541.

Research output: Contribution to journalArticle

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T1 - Complete mapping of disulfide linkages for etanercept products by multi-enzyme digestion coupled with LC-MS/MS using multi-fragmentations including CID and ETD

AU - Huang, Li Juan

AU - Chiang, Chia Wang

AU - Chen, Shun Li

AU - Wei, Shih Yao

AU - Chen, Shu-Hui

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