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- W2297595113 abstract "In order to develop monoclonal antibodies(McAbs) against VT1B of enterohemorrhagic Escherichia coli(EHEC),a VT1B fragment was amplified from E.coli EDL933 by PCR and digested with the endonucleases BamHⅠ and XhoⅠ.After being purified,the fragment was inserted into the expression vector pGEX-6P-1.The recombinant plasmid was named pGEX-VT1B,and transformed into E.coli BL21.The recombinant protein was efficiently expressed in form of inclusion bodies,and made up to 27% of total cell proteins.The BALB/c mice were immunized with the purified recombinant protein to prepare monoclonal antibodies and three strains McAbs 1G11,2H8 and 1B10 were screened from the immunized mice.Western-blotting showed that all McAbs could react specifically against VT1B rather than the GST and were classified into IgG1,IgG2a and IgG2b respectively with the chromosome number of over 100.The indirect ELISA showed that the titer of ascites was over 256000 each and the affinity constant was 0.54×108M-1,0.95×108M-1 and 0.33×107M-1,respectively.The additional ELISA indicated that 1G11,2H8 and 1B10 recognized the same antigenic epitope.The preparation of 3 McAbs provided a basis for detecting Vero toxinⅠ." @default.
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- W2297595113 date "2010-01-01" @default.
- W2297595113 modified "2023-09-22" @default.
- W2297595113 title "Recombinant expression of VT1B gene of enterohemorrhagic Escherichia coli and the preparation of monoclonal antibodies against the expressed recombinant protein" @default.
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