Epitope bias in antibody affinity assays arising from charge-dependent orientation in carbodiimide-mediated protein immobilization using β₂-microglobulin as a model antigen.

Authors

Xiaolan Mai, Linlin Xu, Fang Cheng, Lingyun Jia, Jun Ren

Published in

Chemical communications (Cambridge, England). Sep 23, 2025. Epub Sep 23, 2025.

Abstract

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.

PMID:
40985080
Bibliographic data and abstract were imported from PubMed on 23 Sep 2025.

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