A RECOMBINANT SINGLE-CHAIN IMMUNOTOXIN COMPOSED OF ANTI-TAC VARIABLE REGIONS AND A TRUNCATED DIPHTHERIA-TOXIN

被引：92

作者：

CHAUDHARY, VK ^{[1
]}

GALLO, MG ^{[1
]}

FITZGERALD, DJ ^{[1
]}

PASTAN, I ^{[1
]}

机构：

[1] NCI,DIV CANC BIOL DIAGNOSIS & CTR,MOLEC BIOL LAB,BETHESDA,MD 20892

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 1990年 / 87卷 / 23期

关键词：

antibody; cancer; interleukin; 2; receptor; leukemia; protein engineering;

D O I：

10.1073/pnas.87.23.9491

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

To kill human or primate cells expressing the p55 subunit of the interleukin 2 receptor, we have constructed a single-chain immunotoxin. DNA sequences encoding the first 388 amino acids of diphtheria toxin (DT) were fused to DNA elements encoding the antigen-binding portion (variable region or Fv) of the anti-Tac monoclonal antibody. The antigen-binding portion consists of 116 amino acids of the heavy-chain variable region connected by a 15-amino acid linker to 106 amino acids of the variable region of the light chain. The single-chain immunotoxin DT388-anti-Tac(Fv) was expressed in Escherichia coli and found in inclusion bodies. The monomeric form was then purified to near homogeneity with a high yield (3-5 mg/liter). Monomeric DT388-anti-Tac(Fv) was highly cytotoxic to cell lines bearing the p55 subunit of the human interleukin 2 receptor but not to cells without this subunit. DT388-anti-Tac(Fv) was also very effective in killing proliferating human T cells produced in a mixed leukocyte reaction.

引用

页码：9491 / 9494

页数：4