Aghaei Afshar Mohammad Javad, Robati Reza, Nejatollahi Foroogh
Recombinant Antibody Laboratory, Department of Immunology, Shiraz University of Medical Sciences, Shiraz, Iran.
Shiraz HIV/AIDS Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Rep Biochem Mol Biol. 2019 Oct;8(3):310-317.
Blocking of gp41 of HIV virus, which is involved in the virus entry has been introduced as an effective strategy against HIV infection. In this study we used phage display technology to select specific single chain antibody (scFv) against gp41 HIV for its application in clinical use.
Single chain antibodies against an epitope located in C- terminal part of gp41 were selected using the panning process which enriched a phage antibody display library of scFv. Following panning, 20 clones were amplified by PCR and fingerprinted. To test the specificity of the selected antibodies phage ELISA was performed.
PCR of the library clones demonstrated the presence of VH-linker-VL inserts. Fingerprinting of the clones showed a diverse library with different patterns. Fingerprinting of selected clones after panning revealed two specific single chain antibodies with frequency of 25% and 20%. These clones were preserved for further investigations. Phage ELISA results showed specificity of the two scFvs against the immunodominant epitope of gp41. The absorbance of the scFv1 and scFv2 were 0.72 and 0.63 while the absorbance of the no peptide were 0.18 and 0.12, respectively.
In this study we successfully selected two specific recombinant antibodies against gp41. These libraries are human antibodies with high affinity and specificity and have the potential to be used for diagnosis and treatment. Further investigations are needed to show the effects of the antibodies in vitro and in vivo.
阻断参与病毒进入过程的HIV病毒糖蛋白41（gp41），已被作为一种抗HIV感染的有效策略引入。在本研究中，我们使用噬菌体展示技术筛选针对HIV gp41的特异性单链抗体（scFv），以用于临床应用。
使用淘选过程从scFv噬菌体抗体展示文库中筛选针对位于gp41 C末端部分表位的单链抗体。淘选后，通过PCR扩增20个克隆并进行指纹分析。为了测试所选抗体的特异性，进行了噬菌体ELISA。
文库克隆的PCR证明存在VH-连接子-VL插入片段。克隆的指纹分析显示文库具有不同的模式。淘选后所选克隆的指纹分析揭示了两种特异性单链抗体，频率分别为25%和20%。这些克隆被保存用于进一步研究。噬菌体ELISA结果显示这两种scFv对gp41的免疫显性表位具有特异性。scFv1和scFv2的吸光度分别为0.72和0.63，而无肽对照的吸光度分别为0.18和0.12。
在本研究中，我们成功筛选出两种针对gp41的特异性重组抗体。这些文库是具有高亲和力和特异性的人源抗体，具有用于诊断和治疗的潜力。需要进一步研究以显示这些抗体在体外和体内的作用。
