Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology · 2004
Generation and characterization of monoclonal antibodies against botulinum neurotoxin type A (BoNT/A).
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Wang Jing-lin, Kang Lin, Jia Hong-li
Аннотация
AIM: To generate the monoclonal antibodies(mAbs) against botulinum neurotoxin type A (BoNT/A).
METHODS: BALB/c mice were intraperitoneally immunized with purified BoNT/A-Hc, and the splenocytes of immunized mice were fused with myeloma cells Sp2/0. Hybridoma cells were screened by indirect ELISA and monoclonal hybridoma cells was obtained using limited dilution.
RESULTS: Three hybridomas, named 4A8, 2F7 and 4F2, producing the mAbs against BoNT/A, were successfully established, and the titer of ascitic mAbs ranged from 1x10(-4) to 1x10(-6). Identification of subclass showed that all the produced mAbs belonged to IgG1. 4A8 and 4F2 were stable in secreting anti-BoNT/A mAbs through three-month continuous culture and showed high specificity to recombinant BoNT/A-Hc and native BoNT/A.
CONCLUSION: Anti-BoNT/A mAbs we generated have high specificity, which laid the foundation for the immunological detection of BoNT/A and clinical treatment of botulism in the future.
METHODS: BALB/c mice were intraperitoneally immunized with purified BoNT/A-Hc, and the splenocytes of immunized mice were fused with myeloma cells Sp2/0. Hybridoma cells were screened by indirect ELISA and monoclonal hybridoma cells was obtained using limited dilution.
RESULTS: Three hybridomas, named 4A8, 2F7 and 4F2, producing the mAbs against BoNT/A, were successfully established, and the titer of ascitic mAbs ranged from 1x10(-4) to 1x10(-6). Identification of subclass showed that all the produced mAbs belonged to IgG1. 4A8 and 4F2 were stable in secreting anti-BoNT/A mAbs through three-month continuous culture and showed high specificity to recombinant BoNT/A-Hc and native BoNT/A.
CONCLUSION: Anti-BoNT/A mAbs we generated have high specificity, which laid the foundation for the immunological detection of BoNT/A and clinical treatment of botulism in the future.