Simplified approach for in-vitro production and purification of cell derived Cancer Antigen 15-3

Abstract

Cancer antigen 15-3 (CA15-3) is a key biomarker, currently used for understanding the onset and prognosis of breast cancer. In present investigation, CA15-3 has been purified from the culture supernatant of breast cancer T47-D cell line with 76% yield and 3350 fold purification. Isolated CA15-3 was analyzed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), immunoblotting (western blotting), chemiluminescence immunoassay (CLIA) and Fourier-transform infrared spectroscopy (FTIR). CA15-3 is a monomeric protein with an apparent molecular mass in between ∼250-350kDa. The FTIR spectroscopy revealed similar profiles of T47-D derived CA15-3 and commercially available CA15-3 protein. With the easy availability of T47-D cell line and a simple purification approach described here will support for the large scale production of CA15-3 to be used for various clinical and diagnostic applications.

Keywords: Breast cancer; Cancer antigen 15-3; T47-D breast cancer cell line.