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Construction of single-chain variable fragment antibodies against MCF-7 breast cancer cells.

Ahmad, Zuhaida Asra and Ali, Abdul Manaf and Subramani, Tamilselvan and Mohamed Alitheen, Noorjahan Banu and Hamid, Muhajir and Noor, A. M. and Yeap, Swee Keong (2013) Construction of single-chain variable fragment antibodies against MCF-7 breast cancer cells. Genetics and Molecular Research, 12 (4). 5547-5559. ISSN 1676-5680

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Official URL: http://www.geneticsmr.com/issue/12/4?page=4

Abstract

A phage display library of single chain variable fragment (scFv) against MCF-7 breast cancer cells was constructed from C3A8 hybridoma cells. RNA from the C3A8 was isolated, cDNA was constructed, and variable heavy and light immunoglobulin chain gene region were amplified using PCR. The variable heavy and light chain gene regions were combined with flexible linker, linked to a pCANTAB 5E phagemid vector and electrophoresed into supE strain of Escherichia coli TG1 cells. Forty-eight clones demonstrated positive binding activity to MCF-7 breast cancer cell membrane fragments and the strongest of 48 clones was selected for analysis. The anti-MCF-7 library evaluated by SfiI and NotI digests demonstrated that anti-MCF-7 scFv antibodies possess individual patterns that should be able to recognize distinct human breast cancer cells. The C3A8 scFv, with an apparent molecular weight of 32 kDa, showed high homology (99%) with single chain antibody against rice stripe virus protein P20. In summary, the anti MCF-7 scFv antibody can be used for pretargeting breast cancer for clinical diagnosis of patients; it also has potential for therapeutic applications.

Item Type:Article
Keyword:C3A8; MCF-7; Recombinant antibody; scFv.
Faculty or Institute:Faculty of Biotechnology and Biomolecular Sciences
Institute of Bioscience
Publisher:Fundacao de Pesquisas Cientificas de Ribeirao Preto
DOI Number:10.4238/2013.November.18.5
Altmetrics:http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.4238/2013.November.18.5
ID Code:28118
Deposited By: Umikalthom Abdullah
Deposited On:28 May 2014 16:08
Last Modified:09 Sep 2015 09:11

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