Protocol to determine antibody affinity and concentration in complex solutions using microfluidic antibody affinity profiling

STAR Protoc. 2023 Mar 17;4(1):102095. doi: 10.1016/j.xpro.2023.102095. Epub 2023 Feb 13.

Abstract

Conventional methods of measuring affinity are limited by artificial immobilization, large sample volumes, and homogeneous solutions. This protocol describes microfluidic antibody affinity profiling on complex human samples in solution to obtain a fingerprint reflecting both affinity and active concentration of the target protein. To illustrate the protocol, we analyze the antibody response in SARS-CoV-2 omicron-naïve samples against different SARS-CoV-2 variants of concern. However, the protocol and the technology are amenable to a broad spectrum of biomedical questions. For complete details on the use and execution of this protocol, please refer to Emmenegger et al. (2022),1 Schneider et al. (2022),2 and Fiedler et al. (2022).3.

Keywords: Antibody; Health Sciences; Immunology; Protein Biochemistry; Proteomics; Surface Plasmon Resonance (SPR).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibody Affinity
  • COVID-19*
  • Humans
  • Microfluidics
  • SARS-CoV-2

Supplementary concepts

  • SARS-CoV-2 variants