A rapid surface plasmon resonance (SPR) biosensor immunoassay for screening of somatotropins in injection preparations

Abstract

The use of growth hormones (recombinant somatotropins (rSTs)) is approved in several countries, e.g. the USA, Brazil and Australia to enhance growth or lactating performances of livestock. Their use in the EU is banned, however, due to the widespread application, the illegal use within the EU cannot be excluded. To screen for rSTs in injection preparations, a biosensor immunoassay (BIA) using surface plasmon resonance (SPR) technology was developed. Compared to existing analysis methods for rSTs, like radio immunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA), this technique provides a rapid (7min) alternative. A direct BIA was compared to an indirect (inhibition) BIA and the performances of several antibodies against (r)STs were compared in the indirect BIA. In the final inhibition assay, using rabbit anti-bovine rST, extracts from several injection preparations were shown to contain bovine rST (rbST). The limit of detection for rbST in the assay is 0.008μgmL−1 which is far below the expected concentrations in injection preparations. Although the cross-reactivities for STs of other species were low, screening of injection preparations for porcine, equine and human ST was feasible through the analysis of less diluted extracts. Tryptic digestion followed by nano-electrospray liquid chromatography–ion trap tandem mass spectrometry (nano-LC–MS/MS) was used to identify STs.