Cross-linked polypeptide fragments of .beta.-galactosidase

Details Cross-linked polypeptide fragments of .beta.-galactosidase Cross-linked polypeptide fragments of .beta.-galactosidase

C - Chemistry, Metallurgy

12

N

C12N 9/38 (2006.01) C12Q 1/34 (2006.01) C12Q 1/37 (2006.01)

Patent

CA 2241782

Formation of an intramolecular cross-link in enzyme donor polypeptide fragments of .beta.-galactosidase, thereby forming a cyclic enzyme donor which is hindered from complementation with an enzyme acceptor fragment to form active of .beta.-galactosidase. The cyclic enzyme donor can be linearized by cleaving to restore complementation ability. Assays in which such cyclic enzyme donors are linearized by specific analytes are disclosed, as well as novel homobifunctional bis-maleimido cross-linking agents of formula (I), wherein R is hydroxy or acetate.

Formation d'une réticulation intramoléculaire dans des fragments polypeptides donneurs d'enzyme de beta -galactosidase, produisant un donneur d'enzyme cyclique pour lequel la complémentation avec un fragment accepteur d'enzyme pour former la partie active de la beta -galactosidase est entravée. Le donneur d'enzyme cyclique peut être linéarisé par clivage, ce qui lui rend sa capacité de complémentation. Sont décrites des techniques dans lesquelles de tels donneurs d'enzyme cycliques sont linéarisés par des analytes spécifiques, ainsi que de nouveaux agents de réticulation bis-maleimido homobifonctionnels de formule (I), dans laquelle R est hydroxy ou acétate.

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