Affiliation:
1. State Research Center of Virology and Biotechnology «Vector», Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, 630559, Koltsovo, Novosibirsk oblast
Abstract
A suggested method for obtaining recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) includes the accumulation of the producer strain biomass enriched with the target product up to 30% of total protein content, its isolation and purification. The later consists of the following stages: ultrasound cell disintegration, washing of inclusion bodies with buffer solutions, GM-CSF solubilization from inclusion bodies by 6 M urea, denaturation-renaturation of protein molecules and purification by chromatography on DEAE-Sepharose and combined chromatography on CM-Sepharose and Q-Sepharose followed by dialysis. The proposed method makes it possible to yield up to 10 mg of the protein preparation from 1 g of wet cells with the purity of 98% and high activity shown on the human erythroleukemia cell line.
granulocyte-macrophage colony-stimulating, GM-CSF, producer strain, cultivation, chromatographic purification.
The work was performed in the framework of the State Assignment «Adjustment of the Technology of Preparative Obtaining and Purification of Recombinant Proteins» (no. 13/18).
Publisher
National Research Center Kurchatov Institute
Subject
Ecology,Applied Microbiology and Biotechnology,Biotechnology
Cited by
7 articles.
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