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Updated: Jul 12, 2026
High Yield Expression of Recombinant Human Proteins with the Transient Transfection of HEK293 Cells in Suspension
Published on: December 28, 2015
A purification method for improving the process yield and quality of recombinant human granulocyte colony-stimulating
Dasari Venkata Krishna Rao1, Mangamoori Lakshmi Narasu, Adibhatla Kali Satya Bhujanga Rao
1Research and Development, Biotechnology Division, Natco Research Centre, Sanath Nagar, Hyderabad-500018, India. nrc@natcopharma.co.in
Abstract:
A purification method employing a process-control strategy was developed for improving the yield of rhG-CSF (recombinant human granulocyte colony-stimulating factor). A purity of >/=99% with an overall yield of 2.18 g/l was achieved in the present study. Analysis of the product during purification indicated that detergents removed 72% of LPS (lipopolysaccharides) and 98% of HCPs (host cell proteins) without removing nucleic acid. Cysteine concentration was a key parameter in protein refolding. The bed height and HETP (height equivalent theoretical plates) value in the SEC (size-exclusion chromatography) column was evaluated and its impact on the resolution was studied. Formulation during SEC was found to be crucial for increasing the product yields with saving of time and process costs. The yield obtained in the present study is nearly four times higher than that reported in the literature. The product obtained was found to be acceptable for toxicological studies.
