Surface plasmon resonance immunoassay for biotin determination on a home-made instrument

摘要

In this paper, a home-made SPR analytical system and a denaturalized bovine serum albumin (dBSA) SPR chip were used to detect biotin based on the method of an inhibition immunoassay. The experimental was compared with a commercial protocol. The conditions of regeneration were optimized/The dBSA SPR chip was fabricated by a self-assembling method and the derivative of biotin was immobilized on the dBSA SPR chip by using a on-line mode. A commercial instrument and a dextran-SPR-chip (CM5) were used to implement the same experiments. Based on the experiments of immune reactions between biotin antibody and biotin derivative, the biotin antibody concentration was optimized at about 5μg/mL in order to obtain highly sensitivity, which was used in the followed inhibition immunoassays. The lowest detection limit for biotin is 0.1μg/mL with a detection range of 0.1-1000 μg/ml by using our home-made instrument and the dBSA-SPR-chip. The detection results of our instrument and dBSA SPR chip are comparable with the commercial protocol. Our home-made instrument and dBSA-SPR-chips can be also applied to detect other small molecules based on the indirect inhibition immunoassay.