CONSTRUCTION, BACTERIAL EXPRESSION AND CHARACTERIZATION OF A BIFUNCTIONAL SINGLE-CHAIN ANTIBODY-PHOSPHATASE FUSION PROTEIN TARGETED TO THE HUMAN ERBB-2 RECEPTOR

被引：134

作者：

WELS, W ^{[1
]}

HARWERTH, IM ^{[1
]}

ZWICKL, M ^{[1
]}

HARDMAN, N ^{[1
]}

GRONER, B ^{[1
]}

HYNES, NE ^{[1
]}

机构：

[1] CIBA GEIGY AG,DEPT MOLEC BIOL,BIOTECHNOL SECT,CH-4000 BASEL,SWITZERLAND

来源：

BIO-TECHNOLOGY | 1992年 / 10卷 / 10期

关键词：

D O I：

10.1038/nbt1092-1128

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

We have constructed genes expressing single-chain antigen binding proteins (scFv) which recognize the human erbB-2 receptor. These genes encode the heavy and light chain variable domains of an erbB-2 receptor specific monoclonal antibody, MAb FRP5, connected by a peptide linker. In order to express a bifunctional molecule, a bacterial alkaline phosphatase gene was fused 3' to the scFv gene. The scFv(FRP5) and scFv(FRP5)-alkaline phosphatase fusion protein (scFv(FRP5)-PhoA) expressed in E. coli specifically recognize the human erbB-2 protein and compete with MAb FRP for binding to the receptor. The bound scFv(FRP5)-PhoA protein can be detected directly on tumor cells using a substrate for alkaline phosphatase, showing that the chimeric protein retains both binding and enzymatic activity.

引用

页码：1128 / 1132

页数：5

共 27 条

[1]

BARTON RL, 1991, DRUGS NEWS PERSPECTI, V4, P73

[2]

BERGER MS, 1988, CANCER RES, V48, P1238

[3] SINGLE-CHAIN ANTIGEN-BINDING PROTEINS [J].

BIRD, RE ;

HARDMAN, KD ;

JACOBSON, JW ;

JOHNSON, S ;

KAUFMAN, BM ;

LEE, SM ;

LEE, T ;

POPE, SH ;

RIORDAN, GS ;

WHITLOW, M .

SCIENCE, 1988, 242 (4877) :423-426

[4] B3(FV)-PE38KDEL, A SINGLE-CHAIN IMMUNOTOXIN THAT CAUSES COMPLETE REGRESSION OF A HUMAN CARCINOMA IN MICE [J].

BRINKMANN, U ;

PAI, LH ;

FITZGERALD, DJ ;

WILLINGHAM, M ;

PASTAN, I .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (19) :8616-8620

[5]

CHANG CN, 1986, GENE, V44, P121

[6] A RECOMBINANT IMMUNOTOXIN CONSISTING OF 2 ANTIBODY VARIABLE DOMAINS FUSED TO PSEUDOMONAS EXOTOXIN [J].

CHAUDHARY, VK ;

QUEEN, C ;

JUNGHANS, RP ;

WALDMANN, TA ;

FITZGERALD, DJ ;

PASTAN, I .

NATURE, 1989, 339 (6223) :394-397

[7] INVIVO TUMOR TARGETING OF A RECOMBINANT SINGLE-CHAIN ANTIGEN-BINDING PROTEIN [J].

COLCHER, D ;

BIRD, R ;

ROSELLI, M ;

HARDMAN, KD ;

JOHNSON, S ;

POPE, S ;

DODD, SW ;

PANTOLIANO, MW ;

MILENIC, DE ;

SCHLOM, J .

JNCI-JOURNAL OF THE NATIONAL CANCER INSTITUTE, 1990, 82 (14) :1191-1197

[8] TYROSINE KINASE RECEPTOR WITH EXTENSIVE HOMOLOGY TO EGF RECEPTOR SHARES CHROMOSOMAL LOCATION WITH NEU ONCOGENE [J].

COUSSENS, L ;

YANGFENG, TL ;

LIAO, YC ;

CHEN, E ;

GRAY, A ;

MCGRATH, J ;

SEEBURG, PH ;

LIBERMANN, TA ;

SCHLESSINGER, J ;

FRANCKE, U ;

LEVINSON, A ;

ULLRICH, A .

SCIENCE, 1985, 230 (4730) :1132-1139

[9] A COMPARISON OF STRATEGIES TO STABILIZE IMMUNOGLOBULIN FV-FRAGMENTS [J].

GLOCKSHUBER, R ;

MALIA, M ;

PFITZINGER, I ;

PLUCKTHUN, A .

BIOCHEMISTRY, 1990, 29 (06) :1362-1367

[10]

GUELTEKIN H, 1988, Analytical Biochemistry, V172, P320

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