Renaturation liquid for recombining chicken alpha interferon inclusion body as well as preparation method and application thereof

Abstract

The invention provides a renaturation liquid for recombining a chicken alpha interferon inclusion body as well as a preparation method and an application thereof. The renaturation liquid mainly comprises arginine, EDTA (Ethylene Diamine Tetraacetic Acid), oxidized glutathione, Tris and glycerol, wherein the concentration of each component is as follows: 0.2-0.8 M of arginine, 0.5-2 mM of EDTA, 0.3-0.9 mM of toxidized glutathione, 60-100 mM of Tris and 15-30% (v/v) of glycerol; and the pH (Potential of Hydrogen) of the renaturation liquid is 8.0-8.5. The renaturation liquid is prepared by dissolving in distilled water and utilizing HCl to adjust the pH value; the renaturation liquid has an application prospect on the aspect of recombining the chicken alpha interferon inclusion body; and the renaturation liquid has a reasonable proportion for components to carry out a three-step washing and purifying and drip sample injection primary renaturation process method on the inclusion body to better separate a bacterial protein with the inclusion body, so that the renaturation rate of the protein is greatly improved.

Description

technical field [0001] The invention relates to a refolding liquid and its preparation method and application, in particular to a refolding liquid for recombining chicken alpha interferon inclusion body and its preparation method and application. Background technique [0002] Interferon (English interferon, abbreviated as IFN) is a kind of highly active and multifunctional cytokine secreted by cells. Its main function is to inhibit virus proliferation and anti-tumor, and improve the body's anti-virus ability. Chicken interferon protein with a molecular weight of 17-34KD has protein properties and is stable to heat. It can be stored for a long time at 2-8°C, and its activity can be preserved for a long time at 20°C. It will be destroyed at 56°C. Interferon in the pH range of 2-10 Not destroyed, easily neutralized by nucleases and inactivated by proteases. Genetically engineered chicken alpha interferon is a non-toxic, harmless, and residue-free biological product, which has ...

AI Technical Summary

Problems solved by technology

The dilution method has a large amount of operating liquid and a small protein concentration, which increases the difficulty of subsequent protein purification and increases the production cost; the dialysis method has high requirements for equipment, which increases the investment in equipment. At the same time, the operation time is longer, which may lead to protein precipitation

At present, the renaturation rate of inclusion bodies is about 15-25%, and most of the valuable target proteins cannot be used because they cannot be renatured.

The reason for the low renaturation rate is mainly due to the lack of some cofactors for protein folding during the renaturation process of recombinant proteins, or the inability to form correct secondary bonds due to improper environment, etc.

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