[Primary characterization and sequence analysis of anti-colorectal cancer phage fusion antibodies].
OBJECTIVE
To identify 5 phage fusion antibodies against colorectal cancer from in vitro immunized phage library and analyze their sequences.


METHODS
Cell ELISA, immunohistochemistry, DNA sequencing and computer analysis were employed.


RESULTS
Five clones of phage antibodies were tested by cell ELISA, and all of them reacted to human colorectal cancer cell lines, human embryo kidney endothelial cell line and some tumor cell lines, but not to mouse-original cell lines. They also reacted weakly to human hepatic cell lines. The binding specificity of the phage antibodies for colorectal cancer cells was confirmed by immunohistochemistry with cultured cells and colorectal carcinoma and colon tissue sections. They reacted to colorectal carcinoma cell lines, human embryo kidney endothelial cell lines and nasopharyngeal carcinoma cell lines. CH273 reacted specifically to colorectal cancer cells in human colorectal carcinoma sections but not to any of the cells in human colon sections. The 5 clones were further analyzed after their DNA sequencing. The sequences of CH723, CH209 and CHA12 were identical. The lengths of CH273, CH205 and CH723 were 732 bp, 366 bp and 723 bp, respectively. The VDJ regions of CH273, CH205 and CH723 belonged to VH3-30-D1-26-JH3-linker-V1-13-JL2, VH1-46-D6-13-JH3 and VH3-30-D1-26-JH3-linker-L2-J kappa 2, respectively.


CONCLUSIONS
Phage antibodies binding to colorectal tissues and cells are confirmed, on which human anti-tumor ScFv and VH fragments may be further developed and applied to clinical therapy.
Zhu
Jian-gao
JG
Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha, 410078, China.
Hu
Jin-yue
JY
Li
Guan-cheng
GC
D000595:Amino Acid Sequence; D000818:Animals; D000912:Antibodies, Neoplasm / Q000235:genetics*; D001435:Bacteriophages / Q000276:immunology*; D001483:Base Sequence; D018797:Cell Cycle Proteins / Q000235:genetics; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000276:immunology* / Q000473:pathology; D006360:Heat-Shock Proteins / Q000235:genetics; D006801:Humans; D007147:Immunoglobulin Light Chains / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000235:genetics; D051379:Mice; D008840:Microfilament Proteins; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000276:immunology; D029702:Schizosaccharomyces pombe Proteins; D017421:Sequence Analysis; D014407:Tumor Cells, Cultured
D000912:Antibodies, Neoplasm; D018797:Cell Cycle Proteins; D006360:Heat-Shock Proteins; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D008840:Microfilament Proteins; D011993:Recombinant Fusion Proteins; C099792:Scf1 protein, S pombe; D029702:Schizosaccharomyces pombe Proteins; C115237:actin interacting protein 1