2011•Zhongguo shengwuzhipinxue zazhiRequires access
Preparation and Purification of Polyclonal Antibody against Recombinant P1 Protein of Mycoplasma pneumoniae
Guizhen Wang
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Abstract
Objective To prepare the polyclonal antibody against recombinant P1 protein of Mycoplasma pneumoniae(Mp) and lay a foundation of preparation of diagnostic kit for Mp.Methods Rabbits were immunized with recombinant P1 protein of Mp and determined for antibody titer of immune sera by ELISA.The antibody was purified by amino sulfate precipitation and ion exchange chromatography,and determined for concentration by ultraviolet spectrophotometry and for purity by SDS-PAGE.The purified antibody was labeled by quantum dots(QDs),determined for immunological activity and analyzed for specificity by direct IFA.Results The titer of antisera of rabbits immunized with recombinant P1 protein was 1 ∶ 25 600.The purified antibody,at a concentration of 3.689 μg / μl,showed a heavy chain band with a relative molecular mass of 58 000 on SDS-PAGE profile.The purified antibody labeled by QDs showed high immunological activity but no specific binding to other common respiratory pathogens.Conclusion The purified IgG against P1 protein of Mp showed high immunological activity and specificity,which might be used for antigenic diagnosis Mp infection in clinic.