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. 2023:2552:409-433.
doi: 10.1007/978-1-0716-2609-2_23.

Analytical Method for Experimental Validation of Computer-Designed Antibody

Aki Tanabe  1   2 , Kouhei Tsumoto  3   4   5
Affiliations

Analytical Method for Experimental Validation of Computer-Designed Antibody

Aki Tanabe et al. Methods Mol Biol. 2023.

Abstract

In the computational design of antibodies, the interaction analysis between target antigen and antibody is an essential process to obtain feedback for validation and optimization of the design. Kinetic and thermodynamic parameters as well as binding affinity (KD) allow for a more detailed evaluation and understanding of the molecular recognition. In this chapter, we summarize the conventional experimental methods which can calculate KD value (ELISA, FP), analyze a binding activity to actual cells (FCM), and evaluate the kinetic and thermodynamic parameters (ITC, SPR, BLI), including high-throughput analysis and a recently developed experimental technique.

Keywords: Affinity; Biolayer interferometry (BLI); ELISA; Experimental validation; Flow cytometry (FCM); Fluorescence polarization (FP); Interaction analysis of antigen–antibody; Kinetic parameters; Microscale thermophoresis (MST); Thermodynamic parameters.

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