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  • Implementation of a SPR immunosensor for the simultaneous detection of the 22K and 20K hGH isoforms in human serum samples.

Implementation of a SPR immunosensor for the simultaneous detection of the 22K and 20K hGH isoforms in human serum samples.

Talanta (2013-08-21)
Elena de Juan-Franco, J M Rodríguez-Frade, M Mellado, Laura M Lechuga
ABSTRACT

We have implemented a Surface Plasmon Resonance (SPR) immunosensor based on a sandwich assay for the simultaneous detection of the two main hGH isoforms, of 22 kDa (22K) and 20 kDa (20K). An oriented-antibody sensor surface specific for both hormone isoforms was assembled by using the biotin-streptavidin system. The immunosensor functionality was checked for the direct detection of the 22K hGH isoform in buffer, which gave high specificity and reproducibility (intra and inter-assay mean coefficients of variation of 8.23% and 9% respectively). The selective determination of the 22K and 20K hGH isoforms in human serum samples in a single assay was possible by using two specific anti-hGH monoclonal antibodies. The detection limit for both hormone isoforms was 0.9 ng mL(-1) and the mean coefficient of variation was below 7.2%. The excellent reproducibility and sensitivity obtained indicate the high performance of this immunosensor for implementing an anti-doping test.

MATERIALS
Product Number
Brand
Product Description

Supelco
Biotin, certified reference material, TraceCERT®
Sigma-Aldrich
Biotin, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
Sigma-Aldrich
Biotin, meets USP testing specifications
Sigma-Aldrich
Biotin, tested according to Ph. Eur.
Sigma-Aldrich
Biotin, ≥99.0% (T)
Biotin, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Biotin, Vetec, reagent grade, ≥99%
Supelco
Biotin, Pharmaceutical Secondary Standard; Certified Reference Material