A Rapid and Simple Approach to Preparation of Monoclonal Antibody Based on DNA Immunization

来源 :Cellular & Molecular Immunology | 被引量 : 0次 | 上传用户:xingnaizheng
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Inoculation with purified specific protein is usually the first step for preparation of monoclonal antibody (mAb).But it is quite difficult to obtain pure proteins especially with natural structures.Here we attempt to replace theprotein inoculation with DNA immunization in the preparation of mAb.The eukaryotic expression vectorspcDNA3-PreS2/S and pVAX-PreS2/S encoding the HBV M protein were constructed and prepared for DNAimmunization.Female BALB/c mice developed a well antibody response to the target antigen after muscleinjection with corresponding plasmids.The mice with effective antibodies induced were used for preparation ofmAb.We found the mice immunized with three administrations of pcDNA3-PreS2/S and boosted byintrasplenic injection with the same plasmid could be exploited for preparation of mAb.And positivehybridoma cell 2D3 that can secrete specific mAb was cloned and analyzed.Our studies demonstrate that geneimmunization may provide a convenient and efficient way to prepare mAbs.Cellular & Molecular Immunology.2004;1(4):295-299. Inoculation with purified specific protein is usually the first step for preparation of monoclonal antibody (mAb). It is quite difficult to obtain pure proteins especially with natural structures. Here we attempt to replace the protein inoculation with DNA immunization in the preparation of mAb. eukaryotic expression vectorspcDNA3-PreS2 / S and pVAX-PreS2 / S encoding the HBV M protein were constructed and prepared for DNAimmunization.Female BALB / c mice developed a well antibody response to the target antigen after muscle injection with corresponding plasmids. mice with effective antibodies induced were used for preparation of mAb. We found the mice immunized with three administrations of pcDNA3-PreS2 / S and boosted byintrasplenic injection with the same plasmid could be exploited for preparation of mAb. And positive hybridoma cell 2D3 that can becrete specific mAb was cloned and analyzed .Our studies demonstrate that geneimmunization may provide a convenient and efficient way to prepare m Abs. Cellular & Molecular Immunology. 2004; 1 (4): 295-299.
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