PRODUCTION OF RECOMBINANT PROTEIN CRM197 IN ESCHERICHIA COLI

被引：3

作者：

Dukhovlinov, I. V. ^{[1
]}

Fedorova, E. A. ^{[1
]}

Bogomolova, E. G. ^{[1
]}

Dobrovolskaya, O. A. ^{[1
]}

Chernyaeva, E. N. ^{[1
]}

Al-Shekhadat, R. I. ^{[1
]}

Simbirtsev, A. S. ^{[1
]}

机构：

[1] Res Inst Highly Pure Biopreparat, Pudozhskaya Str 7, St Petersburg 197110, Russia

来源：

INFEKTSIYA I IMMUNITET | 2015年 / 5卷 / 01期

关键词：

recombinant protein; CRM197; adjuvant; conjugate vaccine; diphtheria toxin; Escherichia coli;

D O I：

10.15789/2220-7619-2015-1-37-44

中图分类号：

R51 [传染病];

学科分类号：

100401 ;

摘要：

The CRM197 is a non-toxic mutant of diphtheria toxin having a single amino acid substitution of a glycine for a glutamic acid in position 52. Being naturally nontoxic, CRM197 is a promising adjuvant and ideal carrier protein for conjugate vaccines. Typically, production of diphtheria toxin and some of the non-toxic proteins are carried out by Corynebacterium diphtheriae. Production of recombinant protein CRM197 in Escherichia coli is advantageous. It is simple, cheap and permits production of the target protein in a short time using a non-patho-genic microorganism. In this study patented high-yield-producing E. coli strain was used. As a part of the study the following steps were taken: protocol adjustment for induction of crm197 gene, production and purification of recombinant CRM197 by ion-exchange, hydrophobic and gel-filtration chromatography. The purity of the final preparation reached 97%.

引用

页码：37 / 44

页数：8

共 25 条

[1]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[2] Beta-glucan-CRM197 conjugates as candidates antifungal vaccines [J].

Bromuro, Carla ;

Romano, Maria ;

Chiani, Paola ;

Berti, Francesco ;

Tontini, Marta ;

Proietti, Daniela ;

Mori, Elena ;

Torosantucci, Antonella ;

Costantino, Paolo ;

Rappuoli, Rino ;

Cassone, Antonio .

VACCINE, 2010, 28 (14) :2615-2623

[3] Safety and Immunogenicity of a 13-Valent Pneumococcal Conjugate Vaccine [J].

Bryant, Kristina A. ;

Block, Stan L. ;

Baker, Sherryl A. ;

Gruber, William C. ;

Scott, Daniel A. .

PEDIATRICS, 2010, 125 (05) :866-875

[4] Glycoconjugate vaccines and immune interference: A review [J].

Dagan, Ron ;

Poolman, Jan ;

Siegrist, Claire-Anne .

VACCINE, 2010, 28 (34) :5513-5523

[5] Compaction agent clarification of microbial lysates [J].

DeWalt, BW ;

Murphy, JC ;

Fox, GE ;

Willson, RC .

PROTEIN EXPRESSION AND PURIFICATION, 2003, 28 (02) :220-223

[6] Persistence of immune responses after a single dose of Novartis meningococcal serogroup A, C, W-135 and Y CRM-197 conjugate vaccine (Menveo®) or Menactra® among healthy adolescents [J].

Gill, Christopher J. ;

Baxter, Roger ;

Anemona, Alessandra ;

Ciavarro, Giuseppe L. ;

Dull, Peter M. .

HUMAN VACCINES, 2010, 6 (11) :881-887

[7]

Hwang KW, 2010, ARCH PHARM RES, V33, P793, DOI 10.1007/s12272-010-0600-z

[8]

Kunami N, 2011, ANTICANCER RES, V31, P2483

[9] Transcytosis of CRM197-grafted polybutylcyanoacrylate nanoparticles for delivering zidovudine across human brain-microvascular endothelial cells [J].

Kuo, Yung-Chih ;

Chung, Chiu-Yen .

COLLOIDS AND SURFACES B-BIOINTERFACES, 2012, 91 :242-249

[10] CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].

LAEMMLI, UK .

NATURE, 1970, 227 (5259) :680-+

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