Preparative electroelution of specific protein antigens from MicoplasmaPneumoniae: use in an enazyme-linked immunosorbent assay (Ellisa) |
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| Institution: | 1. The Affiliated Children''s Hospital of Kunming Medical University, Kunming 650228, PR China;2. Institute of Medical Biology, Chinese Academy of Medical Sciences, and Peking Union Medical College, Kunming 650118, PR China;3. Institute of Pediatric Disease Research in Yunnan, Kunming 650228, PR China;4. Yunnan Key Laboratory of Vaccine Research & Development on Severe Infectious Diseases, Kunming 650118, PR China |
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| Abstract: | A rapid, simple and preparative method is described for the recovery of the seven highest molecular weight proteins (HMWP) from Mycoplasma pneumoniae membrane. The yield of proteins obtained was approximately 90%. The method involved the separation of M. pneumoniae proteins by socium dodecyl sulphate polycrylamide gel electrophoresis (SDS-PAGE), followed by electroelution of HMWP.These eluted antigens were used in an ELISA to measure IgG antibodies in sera 9 blood donors and 9 patients with M. pneumoniae infection.The specificity fo M. pneumoniae HMWP was examined by competition ELISA and immunoblotting with different mycoplasma species encountered in teh respiratory tract. |
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