Generation of antibodies against disintegrin and cysteine-rich domains by DNA immunization: An approach to neutralize snake venom-induced haemorrhage

作     者：Sidgi Syed Anwer Abdo Hasson 

作者机构：Department of Microbiology and ImmunologyCollege of Medicine and Health SciencesSultan Qaboos UniversityP.O.Box 35MuscatOman 

出 版 物：《Asian Pacific Journal of Tropical Biomedicine》 (亚太热带生物医学杂志（英文版）)

年 卷 期：2017年第7卷第3期

页      面：198-207页

核心收录：

学科分类：1001[医学-基础医学(可授医学、理学学位)] 100102[医学-免疫学] 10[医学] 

基　　金：Supported by the Wellcome Trust,UK(RAH,Grant No.061325) the University of Science and Technology,Yemen the Gunter Trust,UK 

主　　题：Snake Antivenoms Echis ocellatus GeneGun DNA-immunization Antibody zymography Neutralization 

摘      要：Objective: To explore whether a DNA immunization approach targeting the major haemorrhage molecule of a prothrombin activator-like metalloproteinase from Echis ocellatus(E. ocellatus) venom could be conceived to inspire antibodies with more prominent specificity and equal adequacy to current conventional antivenoms ***: The isolated DNA Eo MP-6 was used as the template for PCR amplification using the Eo DC-2-specific forward and reverse primers. A PCR product of approximately700 bp was obtained and cloned into p Sec Tag-B expression vector where anti-Eo DC-2antibodies were generated and analysed for their efficacy to neutralise local haemorrhage in vitro and in ***: Our results suggest that the generated anti-Eo DC-2 showed a remarkable efficacy by(a) interfering with the interaction of the recombinant disintegrin Eo DC-2 isolated from the E. ocellatus as well as other viper species to the a2b1-integrins on platelets;(b) complete inhibition of the catalytic site of the metalloproteinase molecules in vitro using an adaptation antibody zymography assay. Furthermore, it has a polyspecific potential and constitutively expressed significant inhibition by cross-reaction and neutralised venom-induced local haemorrhage exerted by different viper species in vivo. The potential characteristic of Eo DC-2 against one part(the non-catalytic domain) as opposed to the whole molecule to neutralise its haemorrhagic activity is of crucial importance as it represents a novel approach with greater immunological specificity and fewer hazards, if any, than conventional systems of antivenom production, by exposure large animals that usually being used for the current antivenom production to a less injurious than expression of the whole molecule containing the catalytic metalloprotease domain. Hence, we report for the first time that our preliminary results hold a promising future for antivenom ***: Antibodies generated against the E. ocellatus ve