Preparation of equine immunoglobulin F(ab′)2 against smallpox and evaluation of its immunoprotective effect

Preprint | 
10.55415/deep-2023-0007.v1
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Bochang Shi#
Key Laboratory of Molecular Biology, Inner Mongolia Medical University, Huhehaote Inner Mongolia 010058, China.
Key Laboratory of Molecular Biology, Inner Mongolia Medical University, Huhehaote Inner Mongolia 010058, China.
Hui Han#
Key Laboratory of Molecular Biology, Inner Mongolia Medical University, Huhehaote Inner Mongolia 010058, China.
Key Laboratory of Molecular Biology, Inner Mongolia Medical University, Huhehaote Inner Mongolia 010058, China.
Lingyun Tan#
Department of Immunology, School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China.
Department of Immunology, School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China.
Yuan Liu
College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu Shanxi 030800, China.
College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu Shanxi 030800, China.
Fang Yan
College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu Shanxi 030800, China.
College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu Shanxi 030800, China.
Bo Li
The fifth medical center of PLA General Hospital, Beijing 100039, China.
The fifth medical center of PLA General Hospital, Beijing 100039, China.
Naxin Zheng
The fifth medical center of PLA General Hospital, Beijing 100039, China.
The fifth medical center of PLA General Hospital, Beijing 100039, China.
Min Li
Inner Mongolia Huaxi Biotechnology Co, Ltd., Inner Mongolia 010010, China++Ximu Bio-technology Co., LTD, Beijing 100010, China.
Inner Mongolia Huaxi Biotechnology Co, Ltd., Inner Mongolia 010010, China++Ximu Bio-technology Co., LTD, Beijing 100010, China.
Liyan Zhao
People's Liberation Army Navy's No.971 Hospital, Qingdao Shandong 266071, China.
People's Liberation Army Navy's No.971 Hospital, Qingdao Shandong 266071, China.
Huabin Li
College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu Shanxi 030800, China.
College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu Shanxi 030800, China.
Tietong Fan
Shanghai Serum Biotechnology Co., Ltd, Shanghai 201418, China
Shanghai Serum Biotechnology Co., Ltd, Shanghai 201418, China
Zhiping Zhang
Shanghai Serum Biotechnology Co., Ltd, Shanghai 201418, China
Shanghai Serum Biotechnology Co., Ltd, Shanghai 201418, China
Xinyu Li
Department of Immunology, School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China.
Department of Immunology, School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China.
Chongyu Tian
College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu Shanxi 030800, China.
College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu Shanxi 030800, China.
Yuanqiang Zheng*
Key Laboratory of Molecular Biology, Inner Mongolia Medical University, Huhehaote Inner Mongolia 010058, China.
Key Laboratory of Molecular Biology, Inner Mongolia Medical University, Huhehaote Inner Mongolia 010058, China.
Wei He*
Department of Immunology, School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China.
Department of Immunology, School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China.
Zhongpeng Zhao*
Department of Immunology, School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China.
Department of Immunology, School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China.

# contributed equally to this work, * Corresponding author

Abstract

Objective: Smallpox is a severe infectious disease caused by the smallpox virus, and the death rate is as high as 30% within 15–20 days post infection. 

Methods: We prepared and tested pepsin-digested F(ab’)2 fragments of serum IgGs from horses. 

Results: Transmission electron microscopy showed that the purified virus conformed to the morphology of VVTT. The titer was above 1.0 × 107 PFU/mL. The purity quotient of purified antigen is more than 90% by HPLC. After purification and cutting, and the yield of the purified product F(ab')2 was about 1.3%, the purity of F(ab')2 was more than 90%, and the neutralizing antibody titer was over 1:3200. F(ab')2 fragments have a good preventive and therapeutic effect in mice at antibody dosages of 5.2 mg/mL and 2.6 mg/mL. Compared with the model group, the viral loads of the drug-treated mice were all suppressed to varying degrees, with the higher dose groups (5.2 and 2.6 mg/mL) showing a 2-3 folds reduction in viral loads. 

Conclusion: The process for producing equine immunoglobulin F(ab')2 against VVTT was established. The prepared horse anti-smallpox immunoglobulin product has a good neutralizing antibody effect on VVTT. The highly purified preparation should be a potential candidate for smallpox treatment.

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Keywords
Smallpox ;  Equine immunoglobulin F(ab')2 fragments ;  Needle-free ;  Immunoprotective
Subject Area
Immunology ;  General Medical Research
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