Open AccessA novel click chemistry-based peptide ELISA protocol: development and technical evaluation
Author(s) -
Lisa Milchram,
Regina Soldo,
Valerie Regele,
Silvia Schönthaler,
Markus Degeorgi,
Sophie Baumgartner,
Elena Kopp,
Andreas Weinhäusel
Publication year - 2022
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2021-0107
Subject(s) - click chemistry , peptide , bovine serum albumin , chemistry , detection limit , conjugated system , protocol (science) , avidin , antibody , chromatography , reproducibility , combinatorial chemistry , computational biology , biochemistry , biotin , biology , immunology , medicine , organic chemistry , alternative medicine , pathology , polymer
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.