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Preparation and Characterization of Porcine a-interferon Monoclonal Antibody

Author: HeYanLing
Tutor: JingZhiZhong
School: Gansu Agricultural University
Course: Preventive Veterinary Medicine
Keywords: Recombinant porcine IFN - alpha Recombinant protein Monoclonal antibody Characterization
CLC: R392
Type: Master's thesis
Year: 2009
Downloads: 68
Quote: 0
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Abstract


IFN-α (interferon alpha, IFN-α) is produced by cells of a glycoprotein having biological activity in the role of a specific inducer , the same species of animal cells can be induced to produce a broad spectrum of antiviral activity . Nearly 100 kinds of disease through animal to humans has been found , many of them for viral diseases , some not yet found the right vaccines and drugs to prevent and treat some of the biological characteristics of the virus has not yet study . IFN - alpha is the strongest interferon antiviral IFN - alpha clinical application , but whether it is medical or veterinary medicine remains extremely limited, so to carry out the development of IFN - alpha monoclonal antibody , anti- virus levels for the evaluation of the body as well as IFN - alpha role mechanism of the foundation , has an important role in the promotion of IFN - alpha in the widely used clinically . In this study, hybridoma technology , recombinant porcine IFN - alpha protein immunization of BALB / c mice were prepared under sterile conditions, immune spleen cells were fused with myeloma cells SP2 / 0 cells by GST and GST - pIFN - alpha double screening and multiple cloning culture, 3 to secrete anti- IFN - alpha monoclonal hybridoma stable cell lines . Subtype identification kit with monoclonal antibodies identified subtypes , to determine a monoclonal antibody IgG2a monoclonal antibody IgG2b . The chromosome count of the hybridoma cell lines that secrete monoclonal antibodies , hybridoma cell lines chromosome average of 92 , while the the chromosome average number of myeloma cell lines only 66 , from a genetic point of view proved hybridoma cell lines ; hybridoma ascites and cell supernatant was detected by ELISA , three hybridomas secreting monoclonal antibody titer in the ascites can be up to 1:105 , and the supernatant titer 1:104 ; monoclonal antibody antigen-binding locus analysis showed that the 3 mAbs of the antigen binding sites ; SDS - PAGE results show that the molecular weight of the heavy chain of the 3 mAbs were 60ku light chain molecular weight above 30ku stability ; hybridoma test, cell culture cloned by three times , the supernatant antibody titers between 1:104 to 1:105 , the ability of the three strains of hybridoma cells secrete monoclonal antibodies is stable , has good genetic stability .

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CLC: > Medicine, health > Basic Medical > Medical Immunology
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