PRODUCTION AND PURIFICATION OF POLYCLONAL ANTIBODY AGAINST F(AB')2 FRAGMENT OF HUMAN IMMUNOGLOBULIN G

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NASIRI HADI; VALEDKARIMI ZAHRA; AGHEBATI MALEKI LEILI; ABDOLALIZADEH JALAL; KAZEMI TOHID; ESPARVARINHA MOJGHAN; MAJIDI JAFAR

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Journal: VETERINARY RESEARCH FORUM Year:2017 | Volume:8 | Issue:4 Page(s): 307-312

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Title

PRODUCTION AND PURIFICATION OF POLYCLONAL ANTIBODY AGAINST F(AB')2 FRAGMENT OF HUMAN IMMUNOGLOBULIN G

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Keywords

F(AB')2 FRAGMENTQ1

IMMUNOGLOBULIN GQ1

PEPSIN DIGESTIONQ2

POLYCLONAL ANTIBODYQ1

PURIFICATIONQ1

Abstract

Antibodies are essential tools of biomedical and biochemical researches. Polyclonal antibodies are produced against different epitopes of antigens. Purified F(ab')2 can be used for animal’s immunization to produce polyclonal antibodies. Human IMMUNOGLOBULIN G (IgG) was purified by ion exchange chromatography method. In all stages verification method of the purified antibodies was sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Purified IgG was digested by pepsin enzyme and F(ab')2 fragment was purified by gel filtration separation method. For production of POLYCLONAL ANTIBODY, rabbit was immunized by purified F(ab')2 and antibody production was investigated by enzyme-linked immunosorbent assay. Purified anti-IgG F(ab')2 was conjugated with fluorescein isothiocyanate. Ion exchange chromatography PURIFICATION yielded 38 mg of human IgG antibody. The results of SDS-PAGE in reduced and non-reduced conditions showed bands with 25-30 kDa molecular weight (MW) and 50-kDa respectively and a distinct band with 150 kDa MW. The results of non-reduced SDS-PAGE for determining the purity of F(ab')2 fragment showed one band in 90 kDa and a band in 150 kDa MW position. PURIFICATION by Ion exchange chromatography method resulted about 12 mg rabbit POLYCLONAL ANTIBODY. Flow cytometry showed generated POLYCLONAL ANTIBODY had an acceptable activity compared to commercial antibody. Taking together, purified IgG F(ab')2 and polyclonal anti-IgG F(ab')2 are useful tools in biomedical and biochemical researches and diagnostic kits.

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APA: Copy

NASIRI, HADI, VALEDKARIMI, ZAHRA, AGHEBATI MALEKI, LEILI, ABDOLALIZADEH, JALAL, KAZEMI, TOHID, ESPARVARINHA, MOJGHAN, & MAJIDI, JAFAR. (2017). PRODUCTION AND PURIFICATION OF POLYCLONAL ANTIBODY AGAINST F(AB')2 FRAGMENT OF HUMAN IMMUNOGLOBULIN G. VETERINARY RESEARCH FORUM, 8(4 ), 307-312. SID. https://sid.ir/paper/263628/en

Vancouver: Copy

NASIRI HADI, VALEDKARIMI ZAHRA, AGHEBATI MALEKI LEILI, ABDOLALIZADEH JALAL, KAZEMI TOHID, ESPARVARINHA MOJGHAN, MAJIDI JAFAR. PRODUCTION AND PURIFICATION OF POLYCLONAL ANTIBODY AGAINST F(AB')2 FRAGMENT OF HUMAN IMMUNOGLOBULIN G. VETERINARY RESEARCH FORUM[Internet]. 2017;8(4 ):307-312. Available from: https://sid.ir/paper/263628/en

IEEE: Copy

HADI NASIRI, ZAHRA VALEDKARIMI, LEILI AGHEBATI MALEKI, JALAL ABDOLALIZADEH, TOHID KAZEMI, MOJGHAN ESPARVARINHA, and JAFAR MAJIDI, “PRODUCTION AND PURIFICATION OF POLYCLONAL ANTIBODY AGAINST F(AB')2 FRAGMENT OF HUMAN IMMUNOGLOBULIN G,” VETERINARY RESEARCH FORUM, vol. 8, no. 4 , pp. 307–312, 2017, [Online]. Available: https://sid.ir/paper/263628/en

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