Recombinant Mouse Anti-C. botulinum HA70 Antibody (NAP83-27-7-6) (CAT#: FAMAB-0038CQ)

Figure 1 Antibody binding to recombinant hemagglutinin-GST fusion peptides of hemagglutinins by ELISA.

Microtiter plates were coated with 5 μg/ml of the hemagglutin-GST fusion peptide of BoNT/A complex (A-HA17, A-HA34, A-HA70) and BoNT/B complex (B-HA17, B-HA34, B-HA70). Antibodies are represented as follows: NAP80-5, blue bar; NAP80-7, magenta bar; NAP80-33, yellow bar; NAP83-27, light green bar; NAP83-71, orange bar; NAP83-97, gray bar.

Scotcher, M. C., Cheng, L. W., Ching, K., McGarvey, J., Hnasko, R., & Stanker, L. (2013). Development and characterization of six monoclonal antibodies to hemagglutinin-70 of Clostridium botulinum and their application in a sandwich ELISA. Monoclonal antibodies in immunodiagnosis and immunotherapy, 32(1), 6-15.

Figure 2 Antibody binding on Western blots to BoNT/A and BoNT/B toxin complex component proteins.

900 kDa BoNT/A complex (even numbered lanes), and BoNT/B complex (odd numbered lanes) separated on 10% SDS-PAGE in the presence of 10 mM DTT, transferred to nitrocellulose membranes and probed with MAbs NAP80-5 (lanes 3,4), NAP80-7 (lanes 5,6), NAP80-33 (lanes 7,8), NAP83-27 (lanes 9,10), NAP83-71 (lanes 11,12), and NAP83-97 (lanes 13-14). Lanes 1,2 show the serotype A and B complex components stained with SilverXpress Silver Stain; lanes15,16 show biotinylated BoNT/A and BoNT/B toxin complexes probed with 125 ng/ml Streptavidin-HRP conjugate in TBST (Zymed). Protein relative molecular weight (rMW) is indicated in kDa. Holo, 150 kDa holotoxin; NTNH, 120 kDa non-toxin non-hemagglutinin; N-106, 106 kDa processed fragment of NTNH; Hc, 100 kDa toxin heavy chain; Lc, 50 kDa toxin light chain; HA52, 52 kDa processed fragment of 70 kDa hemagglutinin; HA34, 34 kDa hemagglutinin; HA20, 20 kDa processed fragment of 70 kDa hemagglutinin; HA17, 17 kDa hemagglutinin; N-14, 14 kDa processed fragment of NTNH.

Scotcher, M. C., Cheng, L. W., Ching, K., McGarvey, J., Hnasko, R., & Stanker, L. (2013). Development and characterization of six monoclonal antibodies to hemagglutinin-70 of Clostridium botulinum and their application in a sandwich ELISA. Monoclonal antibodies in immunodiagnosis and immunotherapy, 32(1), 6-15.

Figure 3 Western blot analysis of PTC, serotype A (lanes 1,3,5) and B (lanes 2,4,6) separated by native polyacrylamide gel electrophoresis.

Blots were probed with MAbs NAP80-33 (lanes 1,2) and NAP83-27 (lanes 3,4). Lanes 5,6 represent PTC serotype A and B, respectively, stained with Coomassie.

Scotcher, M. C., Cheng, L. W., Ching, K., McGarvey, J., Hnasko, R., & Stanker, L. (2013). Development and characterization of six monoclonal antibodies to hemagglutinin-70 of Clostridium botulinum and their application in a sandwich ELISA. Monoclonal antibodies in immunodiagnosis and immunotherapy, 32(1), 6-15.

Figure 4 Antibody binding to BoNT/A and BoNT/B 900 kDa neurotoxin complexes by direct binding ELISA.

Microtiter plates coated with 100 mL of a 300 ng/ml solution of BoNT/A toxin complex (♦) or BoNT/Btoxin complex (□). Data represent the average of three replicates – one standard error.

Scotcher, M. C., Cheng, L. W., Ching, K., McGarvey, J., Hnasko, R., & Stanker, L. (2013). Development and characterization of six monoclonal antibodies to hemagglutinin-70 of Clostridium botulinum and their application in a sandwich ELISA. Monoclonal antibodies in immunodiagnosis and immunotherapy, 32(1), 6-15.

Figure 5 Capture of biotin-labeled 900 kDa BoNT/A and BoNT/B toxin complexes by anti-HA70 MAbs.

Biotinylated BoNT/A toxin complex (gray bars) and BoNT/B toxin complex (black bars) were added (100 μL/well of a 600 ng/ml solution) to microtiter wells coated with MAbs F1-2 and MCS6-27 and the six anti-HA70 MAbs. Data are presented as the average of two samples + one standard error.

Scotcher, M. C., Cheng, L. W., Ching, K., McGarvey, J., Hnasko, R., & Stanker, L. (2013). Development and characterization of six monoclonal antibodies to hemagglutinin-70 of Clostridium botulinum and their application in a sandwich ELISA. Monoclonal antibodies in immunodiagnosis and immunotherapy, 32(1), 6-15.