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

Appl Biochem Biotechnol. 2010 Nov;162(5):1435-45. doi: 10.1007/s12010-010-8909-0. Epub 2010 Jan 30.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Neoplasm / immunology*
  • Antibody Specificity / immunology
  • Antigens, Neoplasm / immunology*
  • Blotting, Western
  • Cell Adhesion Molecules / immunology*
  • Colorectal Neoplasms / blood
  • Colorectal Neoplasms / immunology*
  • Drosophila melanogaster / cytology*
  • Electrophoresis, Polyacrylamide Gel
  • Epithelial Cell Adhesion Molecule
  • Female
  • Glycosylation
  • HCT116 Cells
  • Humans
  • Immunoglobulin Fc Fragments / immunology*
  • Mice
  • Mice, Inbred BALB C
  • Polysaccharides / analysis
  • Polysaccharides / chemistry
  • Recombinant Fusion Proteins / immunology*
  • Transformation, Genetic*

Substances

  • Antibodies, Neoplasm
  • Antigens, Neoplasm
  • Cell Adhesion Molecules
  • Epithelial Cell Adhesion Molecule
  • Immunoglobulin Fc Fragments
  • Polysaccharides
  • Recombinant Fusion Proteins