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

Publikation: Beitrag in FachzeitschriftArtikelBegutachtung

Abstract

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.

OriginalspracheEnglisch
Seiten (von - bis)134-142
Seitenumfang9
FachzeitschriftBioTechniques
Jahrgang72
Ausgabenummer4
DOIs
PublikationsstatusVeröffentlicht - Apr. 2022
Extern publiziertJa

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