SEA-scFv as a bifunctional antibody: Construction of a bacterial expression system and its functional analysis

被引：17

作者：

Sakurai, N

Kudo, T

Suzuki, M

Tsumoto, K

Takemura, S

Kodama, H

Ebara, S

Teramae, A

Katayose, Y

Shinoda, M

Kurokawa, T

Hinoda, Y

Imai, K

Matsuno, S

Kumagai, I

机构：

[1] Tohoku Univ, Inst Dev Aging & Canc, Cell Resource Ctr Biomed Res, Aoba Ku, Sendai, Miyagi 9808575, Japan

[2] Tohoku Univ, Sch Med, Dept Surg 1, Sendai, Miyagi 9808575, Japan

[3] Tohoku Univ, Grad Sch Engn, Dept Biomol Engn, Sendai, Miyagi 9808575, Japan

[4] Tohoku Univ, Coll Med Sci, Dept Med Technol, Sendai, Miyagi 980, Japan

[5] Sapporo Med Univ, Sch Med, Dept Internal Med, Sapporo, Hokkaido, Japan

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 1999年 / 256卷 / 01期

关键词：

D O I：

10.1006/bbrc.1999.0263

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A SEA-antibody single chain Fv (SEA-scFv) fusion protein was produced by bacterial expression system in this study. SEA-scFv has both staphylococcal enterotoxin A (SEA) effects and antibody activity directed at the epithelial mucin core protein MUC1, a cancer associated antigen. It was expressed mostly in the cytoplasm as an insoluble form. The gene product was solubilized by guanidine hydrochloride, refolded by conventional dilution method, and purified using metal-chelating chromatography. The resulting SEA-scFv fusion protein preparation was found to react with MUC1 and MHC class II antigens and had the ability to enhance cytotoxicity of lymphokine activated killer cells with a T cell phenotype against a human bile duct carcinoma cell line, TFK-1, expressing MUC1. This genetically engineered SEA-scFv fusion protein promises to be an important reagent for cancer immunotherapy. (C) 1999 Academic Press.

引用

页码：223 / 230

页数：8

共 42 条

[1] CHARACTERIZATION OF 2 DISTINCT MHC CLASS-II BINDING-SITES IN THE SUPERANTIGEN STAPHYLOCOCCAL-ENTEROTOXIN-A [J].