THE GLP LARGE-SCALE PREPARATION OF IMMUNOTOXINS CONTAINING DEGLYCOSYLATED RICIN-A CHAIN AND A HINDERED DISULFIDE BOND

被引：44

作者：

GHETIE, V

THORPE, P

GHETIE, MA

KNOWLES, P

UHR, JW

VITETTA, ES

机构：

[1] UNIV TEXAS,SW MED CTR,DEPT MICROBIOL,5323 HARRY HINES BLVD,DALLAS,TX 75235

[2] UNIV TEXAS,SW MED CTR,DEPT PHARMACOL,DALLAS,TX 75235

[3] UNIV TEXAS,SW MED CTR,CTR CANC IMMUNOL,DALLAS,TX 75235

[4] IMPERIAL CANC RES FUND,DRUG TARGETING LAB,LONDON WC2A 3PX,ENGLAND

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 1991年 / 142卷 / 02期

关键词：

MONOCLONAL ANTIBODY; DEGLYCOSYLATED RICIN-A CHAIN; IMMUNOTOXIN; GOOD LABORATORY PRACTICE; LARGE SCALE;

D O I：

10.1016/0022-1759(91)90110-2

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The large scale preparation of two second generation immunotoxins containing murine monoclonal antibodies and deglycosylated ricin A chain is described. The procedure for the preparation of immunotoxins consists of the derivatization of antibody with SMPT and reduction of dgA with DTT followed by their reaction to establish a hindered interchain disulfide bond. The purification of the immunotoxin includes affinity chromatography on Blue-Sepharose to remove the free antibody and gel filtration on Sephacryl S-200HR to remove any high molecular weight material and free dgA. The two immunotoxins were prepared by GLP procedures and tested for yield, composition, purity, sterility and biological activity.

引用

页码：223 / 230

页数：8

共 10 条

[1] PROTEIN THIOLATION AND REVERSIBLE PROTEIN-PROTEIN CONJUGATION - N-SUCCINIMIDYL 3-(2-PYRIDYLDITHIO)PROPIONATE, A NEW HETEROBIFUNCTIONAL REAGENT [J].