Cao J, Liang M, Guo K
Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100052, China.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2000 Dec;14(4):313-6.
Development of recombinant human monoclonal antibody to hepatitis A virus as a emergent measure for prevention of hepatitis A virus infection.
Human neutralizing monoclonal antibody Fab fragments to HAV have been developed by using phage display technique. The heavy and light chains of human IgG Fab genes were amplified from a HAV patient in convalescent stage. The combinatorial phage antibody library was established by inserting both heavy and light chains of Fab genes into phage mid-vector pComb3 and followed by help phage infection after 4 rounds of panning with purified HAV as coated antigen.
The human Fab fragments to HAV were selected and expressed in bacteria.
The specific binding of Fab antibodies to HAV were demonstrated by their reaction with HAV antigen in ELISA. These results provide the basis for further development of a neutralizing recombinant human whole IgG molecule and hold promise for future use in the prophylaxis of HAV infection.
开发重组人甲型肝炎病毒单克隆抗体作为预防甲型肝炎病毒感染的紧急措施。
利用噬菌体展示技术开发了针对甲型肝炎病毒的人中和单克隆抗体Fab片段。从一名甲型肝炎恢复期患者中扩增出人IgG Fab基因的重链和轻链。通过将Fab基因的重链和轻链插入噬菌体中载体pComb3中,建立组合噬菌体抗体文库,并用纯化的甲型肝炎病毒作为包被抗原进行4轮淘选后,再用辅助噬菌体感染。
筛选出针对甲型肝炎病毒的人Fab片段并在细菌中表达。
通过ELISA中Fab抗体与甲型肝炎病毒抗原的反应,证明了Fab抗体与甲型肝炎病毒的特异性结合。这些结果为进一步开发中和重组人全IgG分子提供了基础,并有望在未来用于预防甲型肝炎病毒感染。