Antibody Design for the Quantification of Photosynthetic Proteins and Their Isoforms

Methods Mol Biol. 2024:2790:405-416. doi: 10.1007/978-1-0716-3790-6_21.

Abstract

Antibodies are a valuable research tool, with uses including detection and quantification of specific proteins. By using peptide fragments to raise antibodies, they can be designed to differentiate between structurally similar proteins, or to bind conserved motifs in divergent proteins. Peptide sequence selection and antibody validation are crucial to ensure reliable results from antibody-based experiments. This chapter describes the steps for the identification of peptide sequences to produce protein- or isoform-specific antibodies using recombinant technologies as well as the subsequent validation of such antibodies. The photosynthetic protein Rubisco activase is used as a case study to explain the various steps involved and key aspects to take into consideration.

Keywords: Antibody design; Antibody production; Antibody validation; Immunoblotting; Monospecific antibody; Protein detection; Protein quantification; Recombinant antibody; Rubisco activase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antibodies* / chemistry
  • Antibodies* / immunology
  • Antibodies* / metabolism
  • Photosynthesis
  • Plant Proteins / metabolism
  • Protein Isoforms*

Substances

  • Protein Isoforms
  • Antibodies
  • Plant Proteins