Generation and characterization of monoclonal antibodies against botulinum neurotoxin type A (BoNT/A).
OBJECTIVE
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.


CONCLUSIONS
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.
Wang
Jing-lin
JL
The Army Center of Microbial Detection and Research, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China. wangjl6481@hotmail.com
Kang
Lin
L
Jia
Hong-li
HL
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000918:Antibody Specificity; D019274:Botulinum Toxins, Type A / Q000032:analysis / Q000276:immunology*; D005260:Female; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D019274:Botulinum Toxins, Type A