[Preparation and primary application of monoclonal antibody against benzylpenicillin].

OBJECTIVE

To develop monoclonal antibody(mAb) against benzylpenicillin and to establish a Sandwich ELISA method for relative quantitative analysis of the allergen which induced penicillin allergy commonly in clinic.

METHODS

Penicillin, as a kind of hapten, was conjugated with carrier protein to form complete antigen and then was used to immunize BALB/c mice. Hybridoma cells secrecting mAb against penicillin were developed. Ascites from the immunized mice were purified by caprylic acid-ammonium sulfate precipitation. The specificity of the purified antibodies was detected and the suitable antibodies were used for establishing Sandwich ELISA.

RESULTS

Nine hybridoma cells stably secrecting mAb were prepared through cell fusion, screening and cloning, and five of these purified mAb had relative high affinity. The double-antibody Sandwich ELISA for relative quantitative analysis of benzylpenicilloyl protein was established with a sensitivity of 870 U/L.The average recovery rate was 107.81% and the average intra- and inter-coefficient of varitation (CV) was 6.7% and 9.3% respectively. The method was applied for relative quantitative analysis of benzylpenicilloyl protein from Group A Streptococcus Preparation.

CONCLUSIONS

Nine hybridoma cell strains stably secreting mAb against benzylpenicillin were obtained. The double-antibody Sandwich ELISA for relative quantitative analysis of benzylpenicilloyl protein was established.