Expression method of Haemocoagulase Acutus (Halase) recombinant protein
US11499163B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Oct 30, 2018 |
| Grant date | Nov 15, 2022 |
| Priority date | — |
| Expiry date | Sep 5, 2041 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N2333/96422
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
The disclosure relates to an expression method of a Haemocoagulase Acutus (Halase) recombinant protein. The method includes the following steps: (1) optimizing a halase gene; (2) performing Polymerase Chain Reaction (PCR) amplification on an optimized halase gene; (3) constructing an Agkis-pMCX expression vector, transforming plasmids to competent cells of an Escherichia coli for amplification, screening in an Amp-resistant manner for positive cloning, sequencing and extracting recombinant plasmids with correct sequencing; (4) transfecting recombinant plasmids to CHO cells; and (5) expressing the recombinant protein and identifying. According to the expression method, a recombinant halase is expressed first using the CHO cells cultured in a serum-free suspension manner; and by utilizing a bioengineering means, the actual production problems of insufficient raw material sources and unstable quality of a snake venom product are solved successfully.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.