A rapid and simple approach to preparation of monoclonal antibody based on DNA immunization.
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 the protein inoculation with DNA immunization in the preparation of mAb. The eukaryotic expression vectors pcDNA3-PreS2/S and pVAX-PreS2/S encoding the HBV M protein were constructed and prepared for DNA immunization. Female BALB/c mice developed a well antibody response to the target antigen after muscle injection with corresponding plasmids. The 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 by intrasplenic injection with the same plasmid could be exploited for preparation of mAb. And positive hybridoma cell 2D3 that can secrete specific mAb was cloned and analyzed. Our studies demonstrate that gene immunization may provide a convenient and efficient way to prepare mAbs.
Ni
Yi
Y
Shanghai Medical College, Fudan University, Shanghai 200032, China.
Ma
Ke
K
Ni
Jing
J
Zheng
Xiujuan
X
Wang
Ying
Y
Xiong
Sidong
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000917:Antibody Formation; D000918:Antibody Specificity; D005260:Female; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000276:immunology; D007114:Immunization; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D019444:Vaccines, DNA; D014759:Viral Envelope Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D006514:Hepatitis B Surface Antigens; D007074:Immunoglobulin G; D019444:Vaccines, DNA; D014759:Viral Envelope Proteins