Production process of human horny cell growth factor-2

Abstract

The production process of human horny cell growth factor-2 includes the steps of: culturing engineering colibacillus carrying expression vector selected from pSE280, PSE380 and pSE420 under the proper expression condition and inserting the encoding sequence of human horny cell growth factor in the polyclonal site of the said expression vector so as to express human horny cell growth factor; and separating and purifying the expressed human horny cell growth factor-2. The production process of the present invention has high expression amount and simple separation and purification steps. The present invention provides also corresponding expression vector and engineering bacterial.

Description

technical field [0001] The invention relates to a production method of human keratinocyte growth factor-2 (Keratinocyte Growth Factor-2, KGF-2), an expression vector and a host cell used in the method. Background technique [0002] Treatment of chronic trauma varies with the severity of the injury. Partial and full-thickness lesions are generally treated with dressings and medical or surgical removal of necrotic tissue known as debridement. Antibiotics are available to prevent infection, and partial-thickness to full-thickness lesions represent the largest group of patients with chronic injuries, most in need of treatment with cytokines such as KGF-2. Patients with full-thickness injuries that extend to muscle, tendon, or bone are at great risk of sepsis and are usually treated surgically. The treatment of chronic trauma has always been a clinically difficult problem, and there is no effective means. There are nearly 3 million patients in the United States every year, and...

AI Technical Summary

Problems solved by technology

[0017] At present, in this field, Escherichia coli expresses KGF-2 mostly in the form of inclusion bodies, and the expression amount accounts for 10-15% of the total protein of the bacteria, which requires denaturation and renaturation means, heavy workload, low yield and high cost

For example, HGS uses the pREP4 plasmid as a vector, and bacterial OD 600 When it grows to 0.4-0.6, it is induced with IPTG. After 3-4 hours of induction, the bacteria are obtained, dissolved in 6M guanidine hydrochloride, and then purified. This process cannot be high in KGF-2 expression, and the purification process is complicated (Ruben; Steven M, et al ., Keratinocyte growth factor-2, US6077692)