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

Autor: Milchram L; Competence Unit Molecular Diagnostics, Center for Health and Bioresources, Austrian Institute of Technology, Giefinggasse 4, Vienna, 1210, Austria., Soldo R; Competence Unit Molecular Diagnostics, Center for Health and Bioresources, Austrian Institute of Technology, Giefinggasse 4, Vienna, 1210, Austria., Regele V; Competence Unit Molecular Diagnostics, Center for Health and Bioresources, Austrian Institute of Technology, Giefinggasse 4, Vienna, 1210, Austria., Schönthaler S; Competence Unit Molecular Diagnostics, Center for Health and Bioresources, Austrian Institute of Technology, Giefinggasse 4, Vienna, 1210, Austria., Degeorgi M; Competence Unit Molecular Diagnostics, Center for Health and Bioresources, Austrian Institute of Technology, Giefinggasse 4, Vienna, 1210, Austria., Baumgartner S; Competence Unit Molecular Diagnostics, Center for Health and Bioresources, Austrian Institute of Technology, Giefinggasse 4, Vienna, 1210, Austria., Kopp E; Competence Unit Molecular Diagnostics, Center for Health and Bioresources, Austrian Institute of Technology, Giefinggasse 4, Vienna, 1210, Austria., Weinhäusel A; Competence Unit Molecular Diagnostics, Center for Health and Bioresources, Austrian Institute of Technology, Giefinggasse 4, Vienna, 1210, Austria.
Jazyk: angličtina
Zdroj: BioTechniques [Biotechniques] 2022 Apr; Vol. 72 (4), pp. 134-142. Date of Electronic Publication: 2022 Mar 02.
DOI: 10.2144/btn-2021-0107
Abstrakt: 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.
Databáze: MEDLINE
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