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Int J Biol Macromol. 2018 Feb;107(Pt B):1456-1462. doi: 10.1016/j.ijbiomac.2017.10.012. Epub 2017 Oct 7.

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

Author information

1
Laboratory of Cell Culture and Native Proteins, Research and Development Division, Yashraj Biotechnology Ltd., Navi Mumbai 400705, Maharashtra, India; Department of Medical Biotechnology, MGM Institute of Health Sciences, Kamothe, Navi Mumbai 410209, Maharashtra, India.
2
Laboratory of Cell Culture and Native Proteins, Research and Development Division, Yashraj Biotechnology Ltd., Navi Mumbai 400705, Maharashtra, India.
3
Department of Molecular-Cellular Biology and Biochemistry, National Institute for Research in Reproductive Health, Jehangir Merwanji Street, Parel, Mumbai 400012, Maharashtra, India.
4
Laboratory of Cell Culture and Native Proteins, Research and Development Division, Yashraj Biotechnology Ltd., Navi Mumbai 400705, Maharashtra, India; Department of Molecular-Cellular Biology and Biochemistry, National Institute for Research in Reproductive Health, Jehangir Merwanji Street, Parel, Mumbai 400012, Maharashtra, India. Electronic address: sham83badgujar@gmail.com.
5
Laboratory of Cell Culture and Native Proteins, Research and Development Division, Yashraj Biotechnology Ltd., Navi Mumbai 400705, Maharashtra, India. Electronic address: manoj.chugh2@gmail.com.

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

PMID:
29017885
DOI:
10.1016/j.ijbiomac.2017.10.012
[Indexed for MEDLINE]

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