Acta Pharmacologica Sinica 2005 December; 26 (12): 1479-1484; doi: 10.1111/j.1745-7254.2005.00210.x

 
Original Article
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Preparation and development of equine hyperimmune globulin F(ab')2 against severe acute respiratory syndrome coronavirus1
 

Jia-hai LU2,8,9, Zhong-min GUO2,9, Wen-yu HAN 3,9, Guo-ling WANG2,9, Ding-mei ZHANG2, Yi-fei WANG4, Sheng-yun SUN4, Qin-he YANG4, Huan-ying ZHENG5, Bing L WONG6, Nan-shan ZHONG7

2Sun Yat-sen University, Guangzhou 510080, China; 3The Agriculture School of Jilin University, Changchun 130062, China; 4Ji-nan University, Guangzhou 510632, China; 5Center for Disease Control and Prevention of Guangdong Province, Guangzhou 510300, China;6Advantek Biologics, Hong Kong, China; 7Guangzhou Institute of Respiratory Diseases, Guangzhou Medical College, Guangzhou 510120, China

 

Aim: The resurgence of severe acute respiratory syndrome (SARS) is still a threat because the causative agent remaining in animal reservoirs is not fully understood, and sporadic cases continue to be reported. Developing high titers of anti-SARS hyperimmune globulin to provide an alternative pathway for emergent future prevention and treatment of SARS.

Methods: SARS coronavirus (CoV)F69 (AY313906) and Z2-Y3 (AY394989) were isolated and identified from 2 different Cantonese onset SARS patients. Immunogen was prepared from SARS-CoV F69 strain. Six health horses were immunized 4 times and serum was collected periodically to measure the profile of specific IgG and neutralizing antibodies using indirect enzyme-linked immunosorbent assay and a microneutralization test. Sera were collected in large amounts at the peak, where IgG was precipitated using ammonium sulphate and subsequently digested with pepsin. The product was then purified using anion-exchange chromatography to obtain F(ab')2 fragments.

Results: The specific IgG and neutralizing antibody titers peaked at approximately week 7 after the first immunization, with a maximum value of 1:14210. The sera collected at the peak were then purified. Fragment of approximately 15 g F(ab')2 was obtained from 1 litre antiserum and the purity was above 90% with the titer of 1:5120, which could neutralize the other strain (SARS-CoV Z2-Y3) as well.


Conclusion:
This research provides a viable strategy for the prevention and treatment of SARS coronavirus infection with equine hyperimmune globulin, with the purpose of combating any resurgence of SARS.

 

Keywords: severe acute respiratory syndrome corona-virus; neutralizing antibodies; hyperimmune globulin; cross protection; F(ab')2 fragments

 
1 Project supported by the LIC Foundation of Hong Kong and the Science Foundation of Guangdong Province (No 2003Z3-E0461).

8 Correspondence to Dr Jia-hai LU.
Phn 86-20-8733-0605. Fax 86-20-8733-2438.
E-mail jiahailu@yahoo.com.cn
9 These authors contributed equally to this work.
Received 2005-06-23     Accepted 2005-08-10

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