Expression and immunogenicity of a recombinant chimeric protein of human colorectal cancer antigen GA733-2 and an Fc antibody fragment in stably transformed Drosophila melanogaster S2 cells.
Human colorectal cancer antigen GA733-2 fused to the immunoglobin Fc fragment (GA733-2-Fc) was expressed in stably transformed Drosophila melanogaster S2 cells, and the immunogenicity of recombinant GA733-2-Fc was investigated in mice. Recombinant GA733-2-Fc was secreted into a culture medium with a molecular mass of approximately 65 kDa. Recombinant GA733-2-Fc was purified to homogeneity using affinity fractionation with Protein A sepharose 4 Fast Flow. Recombinant GA733-2-Fc proteins elicited production of specific antibodies against recombinant GA733-2 by immunization through an intraperitoneal route. Recombinant GA733-2-Fc-induced antibodies showed a binding activity to human colorectal carcinoma HCT-116 cells. Secretory recombinant GA733-2-Fc from Drosophila S2 cell systems can be used as an effective experimental antigen for research in cancer vaccine development.
Seok
Yeon Ju
YJ
Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University, Suwon 446-701, Korea.
Kim
Kyung Il
KI
Yoo
Ki Hyun
KH
Hwang-Bo
Jeon
J
Lee
Hyun Ho
HH
Shon
Dong Hwa
DH
Ko
Ki Sung
KS
Kang
Hyung Sik
HS
Lee
Kyung Jin
KJ
Oh
Doo-Byoung
DB
Joung
Young Hee
YH
Chung
In Sik
IS
D000818:Animals; D000912:Antibodies, Neoplasm / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D000951:Antigens, Neoplasm / Q000276:immunology*; D015153:Blotting, Western; D015815:Cell Adhesion Molecules / Q000276:immunology*; D015179:Colorectal Neoplasms / Q000097:blood / Q000276:immunology*; D004331:Drosophila melanogaster / Q000166:cytology*; D004591:Electrophoresis, Polyacrylamide Gel; D000071858:Epithelial Cell Adhesion Molecule; D005260:Female; D006031:Glycosylation; D045325:HCT116 Cells; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D011134:Polysaccharides / Q000032:analysis / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D014170:Transformation, Genetic
D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; D000071858:Epithelial Cell Adhesion Molecule; D007141:Immunoglobulin Fc Fragments; D011134:Polysaccharides; D011993:Recombinant Fusion Proteins