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mljgoums 2019, 13(1): 6-11 |
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Soleimani S, Kamalzadeh M, Lotfi M. Preparation and Characterization of a Neutralizing Monoclonal Antibody against Poliovirus Type 1 (Mahoney Strain). mljgoums 2019; 13 (1) :6-11
URL: http://mlj.goums.ac.ir/article-1-1146-en.html
URL: http://mlj.goums.ac.ir/article-1-1146-en.html
1- Razi Vaccine & Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), P.O. Box 31975-148, Karaj, Iran , sina.soleimani@gmail.com
2- Razi Vaccine & Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), P.O. Box 31975-148, Karaj, Iran
2- Razi Vaccine & Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), P.O. Box 31975-148, Karaj, Iran
Abstract: (6382 Views)
ABSTRACT
Background and Objectives: Poliomyelitis remains a major public health problem in developing countries, which signify the need for extensive diagnostic and prevention research. The aim of the present study was to design monoclonal antibodies (MAbs) against poliovirus type I with biomedical, diagnostic and therapeutic applications.
Methods: B-cells were isolated from a mouse challenged with polio antigen injection. The B-cell were fused with myeloma tumor cells. After evaluation and screening of approximately 250 hybridoma colons by ELISA, 35 colons with the highest antibody titer and no cross-reactivity were selected and subsequently cloned by limiting dilution. Finally, three colons capable of secreting MAbs against epitopes of poliovirus type I were used for MAb production. Next, the MAbs were characterized by antibody assays, isotyping, epitope analysis (western blot), cross-reactivity test, stability test, sterility test and mycoplasma test.
Results: The results indicated that the MAbs were of IgG1 kappa chain, had good stability and no cross-reactivity. In western blot, a band at 26 kDa which is associated to VP3 neutralization protein was observed.
Conclusion: These serotype-specific MAbs can be potentially used for identification of type I poliovirus for research, diagnostic and prevention purposes.
Keywords: Monoclonal antibody, Hybridoma, Poliomyelitis, Poliovirus.
Background and Objectives: Poliomyelitis remains a major public health problem in developing countries, which signify the need for extensive diagnostic and prevention research. The aim of the present study was to design monoclonal antibodies (MAbs) against poliovirus type I with biomedical, diagnostic and therapeutic applications.
Methods: B-cells were isolated from a mouse challenged with polio antigen injection. The B-cell were fused with myeloma tumor cells. After evaluation and screening of approximately 250 hybridoma colons by ELISA, 35 colons with the highest antibody titer and no cross-reactivity were selected and subsequently cloned by limiting dilution. Finally, three colons capable of secreting MAbs against epitopes of poliovirus type I were used for MAb production. Next, the MAbs were characterized by antibody assays, isotyping, epitope analysis (western blot), cross-reactivity test, stability test, sterility test and mycoplasma test.
Results: The results indicated that the MAbs were of IgG1 kappa chain, had good stability and no cross-reactivity. In western blot, a band at 26 kDa which is associated to VP3 neutralization protein was observed.
Conclusion: These serotype-specific MAbs can be potentially used for identification of type I poliovirus for research, diagnostic and prevention purposes.
Keywords: Monoclonal antibody, Hybridoma, Poliomyelitis, Poliovirus.
Research Article: Original Paper |
Received: 2018/11/24 | Accepted: 2018/11/24 | Published: 2018/11/24 | ePublished: 2018/11/24
Received: 2018/11/24 | Accepted: 2018/11/24 | Published: 2018/11/24 | ePublished: 2018/11/24
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