Label-free quantification of antibody–oligonucleotide conjugates with varying stoichiometries using capillary zone electrophoresis

Mo Zhou; Xiaoxia Zhang; Qinglin Xia; Dongke Liu; Hongxu Chen; Yanli Wen; Lele Wang; Linjie Guo; Jiang Li; Lihua Wang

doi:10.1039/D5AN00214A

Label-free quantification of antibody–oligonucleotide conjugates with varying stoichiometries using capillary zone electrophoresis†

Mo Zhou,‡^ab Xiaoxia Zhang,‡^e Qinglin Xia,^ab Dongke Liu,^e Hongxu Chen,^e Yanli Wen,^d Lele Wang,^d Linjie Guo,

*^c Jiang Li^c and Lihua Wang*^c

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* Corresponding authors

^a CAS Key Laboratory of Interfacial Physics and Technology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800, China

^b University of Chinese Academy of Sciences, Beijing 100049, China

^c Institute of Materiobiology, College of Sciences, Shanghai University, Shanghai 200444, China
E-mail: guolinjie@shu.edu.cn, wanglihua@shu.edu.cn

^d Key Laboratory of Bioanalysis and Metrology, State Administration for Market Regulation, Shanghai Institute of Measurement and Testing Technology, Shanghai 201203, China

^e SCIEX, Shanghai, China

Abstract

We report a label-free capillary zone electrophoresis platform enabling quantitative analysis of antibody–oligonucleotide conjugate (AOC) heterogeneity with single-stranded oligonucleotide (21 nt) resolution. This methodology resolves free antibodies, oligonucleotides, and AOC subpopulations while precisely evaluating conjugation efficiency and oligonucleotide-to-antibody ratio (OAR) heterogeneity across three synthetic strategies.

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Label-free quantification of antibody–oligonucleotide conjugates with varying stoichiometries using capillary zone electrophoresis†

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Label-free quantification of antibody–oligonucleotide conjugates with varying stoichiometries using capillary zone electrophoresis

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