title: Implementation of a SPR immunosensor for the simultaneous detection of the 22K and 20K hGH isoforms in human serum samples.
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.
authors: Nanobiosensors and Bioanalytical Applications Group, Research Center on Nanoscience and Nanotechnology (CSIC) and CIBER-BBN, 08193 Bellaterra, Barcelona, Spain. edjuanf@gmail.com
authors: Elena
authors: E
authors: de Juan-Franco
authors: J M
authors: JM
authors: Rodríguez-Frade
authors: M
authors: M
authors: Mellado
authors: Laura M
authors: LM
authors: Lechuga
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D015374:Biosensing Techniques; D001710:Biotin / Q000737:chemistry; D019382:Human Growth Hormone / Q000097:blood* / Q000276:immunology; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry; D008297:Male; D020033:Protein Isoforms / Q000097:blood / Q000276:immunology; D015203:Reproducibility of Results; D019809:Streptavidin / Q000737:chemistry; D020349:Surface Plasmon Resonance
chemical_list: D000911:Antibodies, Monoclonal; D055767:Immobilized Proteins; D020033:Protein Isoforms; D019382:Human Growth Hormone; D001710:Biotin; D019809:Streptavidin
keywords: Immunosensor; Isoforms; SPR; Serum; Site-directed immobilization; hGH