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Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope
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Epitope bias in antibody affinity assays arising from charge-dependent orientation in carbodiimide-mediated protein

Xiaolan Mai1, Linlin Xu1, Fang Cheng2

  • 1MOE Key Laboratory of Bio-Intelligent Manufacturing and School of Bioengineering, Dalian University of Technology, Dalian, 116024, China. renjun@dlut.edu.cn.

Chemical Communications (Cambridge, England)
|September 23, 2025
PubMed

Carbodiimide reactions are widely employed for protein immobilization. Although coupling to primary amines is theoretically random, the charge on carbodiimide reactive intermediates can induce a preferential orientation of proteins during immobilization. The phenomenon of epitope bias has been observed during the screening of high-affinity antibodies using such immobilized proteins.

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