Related Experiment Video
Updated: Aug 23, 2026
Colorectal Cancer Cell Surface Protein Profiling Using an Antibody Microarray and Fluorescence Multiplexing
Published on: September 25, 2011
[Purification of colorectal cancer-associated antigen by immuno-affinity chromatography]
Yong Yang1, Yu Li, Xian-hui Wang
1Department of Cell Biology, Fourth Military Medical University, Xi'an 710032, China. fmmu_yy@163.com
Aim:
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.
Conclusion:
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.
