Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology · 2007
[Preparation and primary application of monoclonal antibody against benzylpenicillin].
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Wu Yu, Meng Shu-Fang, Hu Chang-Qin, Jin Shao-Hong
Аннотация
AIM: 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.
CONCLUSION: 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.
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.
CONCLUSION: 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.