Scalable production of recombinant three-finger proteins: from inclusion bodies to high quality molecular probes.
Microb Cell Fact
; 23(1): 48, 2024 Feb 12.
Article
in En
| MEDLINE
| ID: mdl-38347541
ABSTRACT
BACKGROUND:
The three-finger proteins are a collection of disulfide bond rich proteins of great biomedical interests. Scalable recombinant expression and purification of bioactive three-finger proteins is quite difficult.RESULTS:
We introduce a working pipeline for expression, purification and validation of disulfide-bond rich three-finger proteins using E. coli as the expression host. With this pipeline, we have successfully obtained highly purified and bioactive recombinant α-Βungarotoxin, k-Bungarotoxin, Hannalgesin, Mambalgin-1, α-Cobratoxin, MTα, Slurp1, Pate B etc. Milligrams to hundreds of milligrams of recombinant three finger proteins were obtained within weeks in the lab. The recombinant proteins showed specificity in binding assay and six of them were crystallized and structurally validated using X-ray diffraction protein crystallography.CONCLUSIONS:
Our pipeline allows refolding and purifying recombinant three finger proteins under optimized conditions and can be scaled up for massive production of three finger proteins. As many three finger proteins have attractive therapeutic or research interests and due to the extremely high quality of the recombinant three finger proteins we obtained, our method provides a competitive alternative to either their native counterparts or chemically synthetic ones and should facilitate related research and applications.Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Inclusion Bodies
/
Escherichia coli
Language:
En
Journal:
Microb Cell Fact
Journal subject:
BIOTECNOLOGIA
/
MICROBIOLOGIA
Year:
2024
Type:
Article
Affiliation country:
United States