Title of article :
Isolation of Specific Human Recombinant Antibodies Against Glycoprotein 41 of HIV
Author/Authors :
Aghaei Afshar, Mohammad Javad Recombinant Antibody Laboratory - Department of Immunology -Shiraz University of Medical Sciences, Iran , Robati, Reza Recombinant Antibody Laboratory - Department of Immunology -Shiraz University of Medical Sciences, Iran , Nejatollahi, Foroogh Recombinant Antibody Laboratory - Department of Immunology -Shiraz University of Medical Sciences, Iran
Pages :
8
From page :
310
To page :
317
Abstract :
Background: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. Methods: 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. Results: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. Conclusions: 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.
Keywords :
HIV virus , scFv , gp41 Panning , Recombinant antibody , Phage display
Journal title :
Reports of Biochemistry and Molecular Biology (RBMB)
Serial Year :
2019
Record number :
2501767
Link To Document :
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