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Procedure for Establishing Downstream Process of Recombinant Platelet Derived Growth Factor Expressed In E. Coli

Polavarapu Srikanth1, S M Jain1, R C Saxena1, and Matadeen Bharti2
1. P G Dept of Zoology, S S L Jain College, Barkatulla University, Bhopal
2. Government Degree College, Jatara Dist. Tikamgarh
Abstract—The clone expressing recombinant platelet derived growth factor was constructed and expressed in E. coli using standard cloning procedure. The upstream process was developed in shake flask level at 5 L capacity and down stream process was tried using preparative HPLC with different mobile phases and by displacement chromatography. The final process selected was using preparative HPLC with mobile phase composition was 20:35:80 (Acetonitrile: Water: 0.01% Trifluroacetic acid). The purified fractions of PDGF were pooled and freeze dried and the final purity was more than 99% by RP-HPLC. 

Index Terms—platelet derived growth factor, purification, chromatography, HPLC

Cite: Polavarapu Srikanth, S M Jain, R C Saxena, and Matadeen Bharti, "Procedure for Establishing Downstream Process of Recombinant Platelet Derived Growth Factor Expressed In E. Coli," International Journal of Life Sciences Biotechnology and Pharma Research, Vol. 2, No. 4, pp. 64-72, October 2013.
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