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Updated: May 12, 2026

Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography
10:50

Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography

Published on: March 9, 2010

Multimodal Physicochemical Characterization of Cross-Linking Activities of Biparatopic Antibodies

Hiroki Akiba1,2, Kohei Shiba3, Kouhei Tsumoto4,5

  • 1Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto, Japan. hakiba@pharm.kyoto-u.ac.jp.

Methods in Molecular Biology (Clifton, N.J.)
|May 10, 2026
PubMed

A biparatopic antibody (BpAb) is a bispecific antibody that recognizes two different epitopes of the same antigen. Many BpAbs have been developed to control the structures of immunocomplexes different from that with the original antibodies. Various binding modes are enabled, including the cross-linking pattern of binding and 1:1-binding, which influence the biological activities of BpAbs. In this chapter, we describe the analytical methods for characterizing cross-linking activities. Taking a development campaign for bivalent asymmetric BpAbs targeting TNFR2 as an example, combined analysis is described using size-exclusion chromatography-multi-angle light scattering, mass photometry, and surface plasmon resonance.

Keywords:
Biepitopic antibodyBiparatopic antibodyImmune complexInteractionLight scatteringMass photometrySize-exclusion chromatographySurface plasmon resonance

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