Preparation and Identification of Polyclonal Antibodies against Salbutamol
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Abstract:
In order to prepare the polyclonal antibody against Salbutamol(SAL), SAL derivatives were prepared by four kinds of reactions, then they were coupled to BSA and OVA, synthesizing complete antigen. The protein concentration was determined by Ultraviolet spectrophotography and the coupling ratios of four immunogens were calculated. New Zealand white rabbits were immunized by four immunogens to develop anti-SAL ant?ibodies. All the antibodies and coating antigenes were screened and then selected the best combination. The polyclonal antiserum was purified by caprylic acid-ammonium sulfate precipitation. Antibody titer and specificity were analyzed by indirect ELISA. It turned out that the study successfully got the coupling of the SAL complete antigen and the coupling ratios of four immunogens were 9:1, 6:1, 6:1, 12:1, respectively. The optimal antibody-coated antigen combination was obtained and antibody titer was 1:64000. The antibody showed high cross-reactivity with clenbuterol(144%) and terbutaline (169%), without notable cross-reactivity with other structural analogues of SAL. This study lays the foundation for the establishment of Salbutamol, Clenbuterol and Terbutaline multi-residue enzyme-linked immunosorbent assay.
