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

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. Method summary Peptides were coupled to a common carrier protein and tested in standard plastics to establish an optimized peptide ELISA protocol. The performance of this approach was compared with well-known coupling systems.

Automated summary

ELISA is the current standard for antibody diagnostics, but peptide-based protocols are rarely available. In this study, the authors investigated an indirect ELISA protocol using peptides conjugated onto a protein carrier based on click chemistry and immobilized in standard plastics, and compared it with the common biotin-avidin system. The results showed a slightly improved limit of detection with the peptide-based approach. This method offers a reliable alternative to standard plastics and plate readers.