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KMID : 0545120190290071165
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Journal of Microbiology and Biotechnology
2019 Volume.29 No. 7 p.1165 ~ p.1176
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Development of an Equine Antitoxin by Immunizing the Halla Horse with the Receptor-Binding Domain of Botulinum Neurotoxin Type A1
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Kim Na-Young
Park Kyung-Eui
Lee Yong-Jin
Kim Yeong-Mun
Hong Sung-Hyun
Son Won-Rak
Hong Sung-Youl
Lee Sae-Hyung
Ahn Hye-Bin
Yang Jae-Hyuk
Seo Jong-Pil
Lim Yoon-Kyu
Yu Chi-Ho
Hur Gyeung-Haeng
Jeong Seong-Tae
Lee Hun-Seok
Song Kyoung
Kang Tae-Jin
Shin Young-Kee
Choi Joon-Seok
Choi Jun-Young
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Abstract
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Botulinum neurotoxins (BoNTs), produced by Clostridium botulinum, are the most toxic substances known. However, the number of currently approved medical countermeasures for these toxins is very limited. Therefore, studies on therapeutic antitoxins are essential to prepare for toxin-related emergencies. Currently, more than 10,000 Halla horses, a crossbreed between the native Jeju and Thoroughbred horses, are being raised in Jeju Island of Korea. They can be used for equine antitoxin experiments and production of hyperimmune serum against BoNT/A1. Instead of the inactivated BoNT/A1 toxoid, Halla horse was immunized with the receptor-binding domain present in the C-terminus of heavy chain of BoNT/A1 (BoNT/A1-HCR) expressed in Escherichia coli. The anti-BoNT/A1-HCR antibody titer increased rapidly by week 4, and this level was maintained for several weeks after boosting immunization. Notably, 20 μL of the week 24 BoNT/A1-HCR(-immunized) equine serum showed an in vitro neutralizing activity of over 8 international unit (IU) of a reference equine antitoxin. Furthermore, 20 μL of equine serum and 100 μg of purified equine F(ab’)2 showed 100% neutralization of 10,000 LD50 in vivo. The results of this study shall contribute towards optimizing antitoxin production for BoNT/A1, which is essential for emergency preparedness and response.
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KEYWORD
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Equine antitoxin, botulinum neurotoxin type A, Halla horse
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