Enzyme amplification by using free enzyme to release enzyme from an immobilized enzyme material
US5190864A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Apr 30, 1990 |
| Grant date | Mar 2, 1993 |
| Priority date | — |
| Expiry date | Apr 30, 2010 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/68
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A method for amplifying enzyme activity is disclosed. Enzyme amplification is achieved by covalently bonding enzyme to a supporting material via a molecular chain which is a substrate for the enzyme, then introducing a small amount of enzyme in the free state to this system, causing release of a large amount of bound enzyme. In an alternative embodiment, complementary enzymatically inactive fragments of an active enzyme, which fragments can recombine to form active enzyme, are covalently attached to separate support materials by a molecular chain material which is a substrate for the active enzyme, and these two fragment-supported conjugates are connected in series. Upon application of free enzyme or free complementary enzyme to one of these fragment-support conjugates, followed by application of the resulting product mixture to the second fragment-support conjugate, a large amount of free enzyme is ultimately produced. In a second alternative embodiment, two different active enzymes are each attached to separate supporting materials by different leashes, in which the leash for the first enzyme only is cleaved in the system by the second enzyme, and the leash for the second enzyme …
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.