Production and purification of polyclonal antibody against F(ab')2 fragment of human immunoglobulin G.
作者： Leili Aghebati-Maleki , Tohid Kazemi , Jalal Abdolalizadeh , Jafar Majidi , Hadi Nasiri
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摘要: Antibodies are essential tools of biomedical and biochemical researches. Polyclonal antibodies produced against different epitopes 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 the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). IgG digested pepsin enzyme fragment filtration separation For production antibody, rabbit immunized antibody investigated enzyme-linked immunosorbent assay. anti-IgG conjugated with fluorescein isothiocyanate. Ion purification yielded 38 mg human antibody. The results SDS-PAGE in reduced non-reduced conditions showed bands 25-30 kDa molecular weight (MW) 50-kDa respectively a distinct band 150 MW. determining purity one 90 MW position. Purification resulted about 12 Flow cytometry generated had an acceptable activity compared commercial Taking together, useful researches diagnostic kits.
参考文章(31)
P.P.A. Marlies Leenaars, Coenraad F.M. Hendriksen, Wim A. de Leeuw, Florina Carat, Philippe Delahaut, René Fischer, Marlies Halder, W. Carey Hanly, Joachim Hartinger, Jann Hau, Erik B. Lindblad, Werner Nicklas, Ingrid M. Outschoorn, Duncan E.S. Stewart-Tull, The Production of Polyclonal Antibodies in Laboratory Animals. The Report and Recommendations of ECVAM Workshop 35. Atla-alternatives To Laboratory Animals. ,vol. 27, pp. 79- 102 ,(1999) , 10.1177/026119299902700106
Jiahai Lu, Zhongmin Guo, Xinghua Pan, Guoling Wang, Dingmei Zhang, Yanbin Li, Bingyan Tan, Liping Ouyang, Xinbing Yu, Passive immunotherapy for influenza A H5N1 virus infection with equine hyperimmune globulin F(ab')2 in mice. Respiratory Research. ,vol. 7, pp. 43- 43 ,(2006) , 10.1186/1465-9921-7-43
Koushan Sineh Sepehr, Behzad Baradaran, Jafar Majidi, Jalal Abdolalizadeh, Leili Aghebati, Fatemeh Zare Shahneh, Development and characterization of monoclonal antibodies against human CD20 in Balb/c mice. Human antibodies. ,vol. 21, pp. 57- 64 ,(2013) , 10.3233/HAB-130263
Leili Aghebati Maleki, Jafar Majidi, Behzad Baradaran, Jalal Abdolalizadeh, Aliakbar Movassaghpour Akbari, Production and characterization of murine monoclonal antibody against synthetic peptide of CD34 Human antibodies. ,vol. 22, pp. 1- 8 ,(2013) , 10.3233/HAB-130265
Behzad Baradaran, Aliakbar Movassaghpour, Leili Aghebati Maleki, Jalal Abdolalizadeh, Jafar Majidi, Generation and Characterization of Anti-CD34 Monoclonal Antibodies that React with Hematopoietic Stem Cells. Cell. ,vol. 16, pp. 361- 366 ,(2014)
Leili Aghebati Maleki, Behzad Baradaran, Jafar Majidi, Mozhdeh Mohammadian, Fatemeh Zare Shahneh, Future prospects of monoclonal antibodies as magic bullets in Immunotherapy Human antibodies. ,vol. 22, pp. 9- 13 ,(2013) , 10.3233/HAB-130266
Duncan Low, Rhona O’Leary, Narahari S. Pujar, Future of antibody purification. Journal of Chromatography B. ,vol. 848, pp. 48- 63 ,(2007) , 10.1016/J.JCHROMB.2006.10.033
Ali ElBakri, Paul N. Nelson, Raed O. Abu Odeh, The state of antibody therapy Human Immunology. ,vol. 71, pp. 1243- 1250 ,(2010) , 10.1016/J.HUMIMM.2010.09.007
Ryutaro Oba, Koji Orihara, Tomoaki Kumagai, Hiroyuki Hirai, Kinya Nagata, Shuichi Hamasaki, Chuwa Tei, Nakamura Masataka, Reevaluation of quantitative flow cytometric analysis for TLR2 on monocytes using F(ab′)2 fragments of monoclonal antibodies Cytometry Part A. ,vol. 79, pp. 247- 255 ,(2011) , 10.1002/CYTO.A.21000
N. S. Lipman, L. R. Jackson, L. J. Trudel, F. Weis-Garcia, Monoclonal versus polyclonal antibodies: distinguishing characteristics, applications, and information resources. Ilar Journal. ,vol. 46, pp. 258- 268 ,(2005) , 10.1093/ILAR.46.3.258