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可溶性活性重组流感嗜血杆菌IgA1蛋白酶在大肠杆菌中的表达。

The expression of soluble and active recombinant Haemophilus influenzae IgA1 protease in E. coli.

作者信息

Long Shinong, Phan Elaine, Vellard Michel C

机构信息

Department of Cellular and Molecular Biology, BioMarin Pharmaceutical Inc., 105 Digital Drive, Novato, CA 94949, USA.

出版信息

J Biomed Biotechnol. 2010;2010:253983. doi: 10.1155/2010/253983. Epub 2010 Nov 30.

Abstract

Immunoglobulin A1 (IgA1) proteases from Haemophilus influenzae are extracellular proteases that specifically cleave the hinge region of human IgA1, the predominant class of immunoglobulin present on mucosal membranes. The IgA1 proteases may have the potential to cleave IgA1 complexes in the kidney and be a therapeutic agent for IgA1 nephropathy (IgAN), a disease characterized by deposition of the IgA1 antibody in the glomerulus. We have screened for the expression of recombinant H. influenzae IgA1 protease by combining various expression plasmids, IgA1 protease constructs, and E. coli strains under multiple conditions. Using the method we have developed, approximately 20-40 mg/L of soluble and active H. influenzae IgA1 protease can be produced from E. coli strain C41(DE3), a significant increase in yield compared to the yield upon expression in H. influenzae or other related bacteria.

摘要

来自流感嗜血杆菌的免疫球蛋白A1(IgA1)蛋白酶是一种细胞外蛋白酶,可特异性切割人IgA1的铰链区,IgA1是粘膜膜上存在的主要免疫球蛋白类别。IgA1蛋白酶可能具有切割肾脏中IgA1复合物的潜力,并且可能成为IgA1肾病(IgAN)的治疗剂,IgA1肾病是一种以IgA1抗体在肾小球中沉积为特征的疾病。我们通过在多种条件下组合各种表达质粒、IgA1蛋白酶构建体和大肠杆菌菌株,筛选了重组流感嗜血杆菌IgA1蛋白酶的表达。使用我们开发的方法,大肠杆菌C41(DE3)菌株可产生约20-40mg/L的可溶性活性流感嗜血杆菌IgA1蛋白酶,与在流感嗜血杆菌或其他相关细菌中表达时的产量相比,产量有显著提高。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7196/2995913/8bf89898909e/JBB2010-253983.001.jpg

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