- 명확한 유전적 배경
- 저비용 및 간단한 배양 조건
- 간단한 형질전환 과정
- 짧은 단백질 발현 기간
- 높은 번식률과 발현 수준
- 선택할 수 있는 다양한 carrier와 fusion tag
- 발현 최적화를 위해 많은 매개변수 변형 가능
- Tag는 절단이 쉬운 효소 절단 부위를 가진 N-terminal과 C-terminal 모두에서 사용 가능
코스모진텍 서비스
해외 서비스
Bacterial Expression
E.coli expression system은 구조가 간단하고 유전적 배경이 명확하여 표적 단백질의 수율이 높습니다. 배양 기간이 짧기 때문에 가장 보편적으로 사용되고 비교적 저렴한 비용에 서비스가 가능합니다. 최근 수십 년간 E.coli 발현 시스템도 지속적으로 개발 및 개선되어 과학 연구자 및 산업용 사용자들이 다수의 재조합 단백질 발현에 집중적으로 사용하고 있습니다.
서비스 장점
활용 분야
- Antigen preparation, Ligand preparation, and expression of cytokines and bacteria proteins
- Purified protein (structure, enzymology, drug discovery)
- Protein therapeutics
Service Process
Process
- Codon optimization;gene synthesis
- Restriction digestion of PCR products; Ligation to expression vector
(e.g. Pcold-SUMO, pGEX-4T-1, pET22b-JT) - Transform TOP10 E.coli competent cells
- Obtain the correct recombinant plasmid
Cusabio Features
Multiple vectors optimization · More options for customers
Optimize multiple vectors at the same time; Select the vector that has the highest yield, which can shorten lead time.
Process
- Transform the recombinant plasmid to host cells
(e.g. BL21(DE3), Rosetta-gami B(DE3) pLysS, C41)
Culture overnight at 37°C - Select single colony for small-scale induced expression; Detect protein expression by SDS-PAGE; Preserve the best colony
- Optimize the expression conditions
Cusabio Features
Multi-conditions optimization · Multi-hosts selection
In the small test, the temperature and IPTG are optimized to obtain the most suitable culture conditions. Multiple hosts are transformed at the same time to select the host bacteria with the highest yield.
Process
- 1–10 L large-scale expression
- Protein purification
Cusabio Features
Multiple purification methods (optional)
Explore different chromatographic conditions including ion exchange, hydrophobic and others by using AKTA, and then determine the optimal purification method.
Process
- Charged Tag removal by restriction digestion
- Free Filter-sterilization
- Free Endotoxin removal
- Free Lyophilization ※ Note : Lyophilization and Filter-sterilization cannot be applied simultaneously
Cusabio Features
Flexible additional services
Customers can flexibly choose from a variety of additional services to their specific needs, e.g. Endotoxin removal, Filter-sterilization, Tag removal, Lyophilization, etc. Some are complimentary, and some require additional charge.
Process
- Testing of purity, concentration, etc.
- QC report is provided
Cusabio Features
Detailed COA report
Detailed product data sheet and COA are provided for each project.