BioTechniques (Apr 2022)

A novel click chemistry-based peptide ELISA protocol: development and technical evaluation

  • Lisa Milchram,
  • Regina Soldo,
  • Valerie Regele,
  • Silvia Schönthaler,
  • Markus Degeorgi,
  • Sophie Baumgartner,
  • Elena Kopp,
  • Andreas Weinhäusel

DOI
https://doi.org/10.2144/btn-2021-0107
Journal volume & issue
Vol. 72, no. 4
pp. 134 – 142

Abstract

Read online

ELISA is the current standard for (auto)antibody diagnostics. Once established, ELISA protocols can be easily adapted for novel antigens; however, peptide-based protocols are rarely available. Herein the authors describe the results of a technical investigation of an indirect ELISA protocol using peptides conjugated onto a protein carrier based on click chemistry and immobilized in standard plastics. The authors compared this approach with the common biotin–avidin system and obtained a slightly improved limit of detection for purified IgG of 25–100 ng/well compared with 25–1000 ng/well. Reproducibility and stability of the methodological approach were conducted for further technical characterization. Indirect ELISA using immunoreactive peptides conjugated to bovine serum albumin offers a reliable method that is complementary to standard plastics and plate readers.

Keywords