[Purification of colorectal cancer-associated antigen by immuno-affinity chromatography].
OBJECTIVE
To purify and characterize the colorectal cancer-associated antigen from cultured colorectal cancer cells.


METHODS
The colorectal cancer cell lines that highly expressed the associated antigen were selected by flow cytometry with five specific monoclonal antibodies (mAbs) CYL1-5. Western blot was used to determine the binding ability of five mAbs to the associated antigens released from colorectal cells lysed with single or triplex-detergent, respectively. The mAb with highest binding ability was employed as the ligand for the immuno-affinity chromatography. The antigens purified through immuno-affinity chromatography were identified by Western blot.


RESULTS
The associated antigens were highly expressed on the colorectal cancer cell lines Hce-8693. The binding ability of mAb CYL-2 to the antigen was higher than that of CYL-1, CYL-3-5. The purified associated antigen binding to mAb CYL-2 was a heterodimer composed of two subunits with relative molecular mass (M(r)) of 60 x 10(3) and 70 x 10(3), respectively.


CONCLUSIONS
The purified associated-antigen binding to mAb CYL-2 was obtained from the colorectal cancer cell line Hce-8693 through immuno-affinity chromatography with mAb CYL-2.
Yang
Yong
Y
Department of Cell Biology, Fourth Military Medical University, Xi'an 710032, China. fmmu_yy@163.com
Li
Yu
Y
Wang
Xian-hui
XH
Shang
Peng
P
Wang
Li
L
Qian
Ai-rong
AR
Yao
Xi-ying
XY
Chen
Zhi-nan
ZN
D000911:Antibodies, Monoclonal / Q000276:immunology; D000951:Antigens, Neoplasm / Q000302:isolation & purification*; D045744:Cell Line, Tumor; D002846:Chromatography, Affinity; D015179:Colorectal Neoplasms / Q000276:immunology* / Q000473:pathology; D006801:Humans
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm