PREPARATION OF WELL-DEFINED PROTEIN CONJUGATES USING ENZYME-ASSISTED REVERSE PROTEOLYSIS

被引：29

作者：

ROSE, K ^{[1
]}

VILASECA, LA ^{[1
]}

WERLEN, R ^{[1
]}

MEUNIER, A ^{[1
]}

FISCH, I ^{[1
]}

JONES, RML ^{[1
]}

OFFORD, RE ^{[1
]}

机构：

[1] UNIV GENEVA,MED CTR,DEPT BIOCHIM MED,1 RUE MICHEL SERVET,CH-1211 GENEVA 4,SWITZERLAND

来源：

BIOCONJUGATE CHEMISTRY | 1991年 / 2卷 / 03期

关键词：

D O I：

10.1021/bc00009a004

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A two-step approach to the production of well-defined protein conjugates is described. In the first step, a linker group, carbohydrazide, having unique reactivity (a hydrazide group) is attached specifically to the carboxyl terminus by using enzyme-catalyzed reverse proteolysis. Since the hydrazide group exists nowhere else on the protein, specificity is assured in a subsequent chemical reaction (formation of a hydrazone bond) of the modified protein with a molecule (chelator, drug, or polypeptide) carrying an aldehyde or keto group. The product is sufficiently stable at neutral pH, no reduction of the hydrazone bond being necessary for the hydrazones described. Protein modification is thus restricted to the carboxyl terminus and a homogeneous product results. With insulin as a model, conditions are described for producing such well-defined conjugates in good yields. The use of other linker groups besides carbohydrazide, and applications of these techniques to antibody fragments, are discussed.

引用

页码：154 / 159

页数：6

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