Sheep Monoclonal Antibodies Prevent Systemic Effects of Botulinum Neurotoxin A1 英文参考文献.docVIP

Toxins 2012, 4, 1565-1581; doi:10.3390/toxins4121565 OPEN ACCESS toxins ISSN 2072-6651 /journal/toxins Article Sheep Monoclonal Antibodies Prevent Systemic Effects of Botulinum Neurotoxin A1 Jean Mukherjee 1, Chase McCann 1, Kwasi Ofori 1, Julia Hill 2, Karen Baldwin 1, Charles B. Shoemaker 1, Peter Harrison 2 and Saul Tzipori 1,* 1 Tufts Cummings School of Veterinary Medicine, North Grafton, MA 01536, USA 2 Bioventix Limited, Farnham Surrey, UK * Author to whom correspondence should be addressed; E-Mail: saul.tzipori@; Tel.: +1-508-839-7955; Fax: +1-508-839-7911. Received: 22 November 2012; in revised form: 13 December 2012 / Accepted: 13 December 2012 / Published: 19 December 2012 Abstract: Botulinum neurotoxin (BoNT) is responsible for causing botulism, a potentially fatal disease characterized by paralysis of skeletal muscle. Existing specific treatments include polyclonal antisera derived from immunized humans or horses. Both preparations have similar drawbacks, including limited supply, risk of adverse effects and batch to batch variation. Here, we describe a panel of six highly protective sheep monoclonal antibodies (SMAbs) derived from sheep immunized with BoNT/A1 toxoid (SMAbs 2G11, 4F7) or BoNT/A1 heavy chain C-terminus (HcC) (SMAbs 1G4, 5E2, 5F7, 16F9) with or without subsequent challenge immunization with BoNT/A1 toxin. Although each SMAb bound BoNT/A1 toxin, differences in specificity for native and recombinant constituents of BoNT/A1 were observed. Structural differences were suggested by pI (5E2 = 8.2; 2G11 = 7.1; 4F7 = 8.8; 1G4 = 7.4; 5F7 = 8.0; 16F9 = 5.1). SMAb protective efficacy vs. 10,000 LD50 BoNT/A1 was evaluated using the mouse lethality assay. Although not protective alone, divalent and trivalent combinations of SMabs, IG4, 5F7 and/or 16F9 were highly protective. Divalent combinations containing 0.5–4 μg/SMAb (1–8 μg total SMAb) were 100% protective agai