Protein expression
Recombinant protein expression and purification Services
E. coli Expression System
E. coli expression system is the earliest developed and most widely used gene expression system. In recent decades, E.coli expression system has been popularly used and developed for producing a wide variety of different types of proteins. Compared with other expression systems, E. coli is a well-established host with the advantages of short culturing time, easy genetic manipulation and low cost media.
Royobiotech Co.,Ltd researchers established a well-developed E. coli expression and purification services platform, which offers theexpression and purification services of a variety of recombinant proteins, with a fast turn-around time and competitive pricing.
Service：
Generation of Recombinant Plasmid
Expression Test in Host
Soluble Expression Test
Small Scale Expression and Purification (2L Cell Culture)
Scale Up Expression and Purification
|
Steps |
Items |
Timeline |
Comments |
|
1 |
Generation of Recombinant Plasmid |
2 weeks |
Multiple expression vector, different affinity tag: His, Trx , GST, SUMO… |
|
2 |
Expression of Recombinant Protein |
1 week |
Multiple expression host |
|
3 |
Soluble Expression Optimization |
1-2 weeks |
Test different induction concentration, time point, temperatures or reconstruct plasmids |
|
4 |
Small Scale Expression and Purification |
1-2 weeks |
Purification of soluble expression or from inclusion body |
|
5 |
Scale Up Expression and Purification |
TBD |
Culture in flask or fermenter |
Remarks:
The service is highly flexible to meet your request. One or more above steps could be separately provided.
Affinity purification, ion exchange purification, hydrophobic beads purification and size exclusion column purification or their combination will be used for protein purification.
Yeast Expression System
Yeast expression system contains the advantages of both prokaryotic and eukaryotic system, such as easy scaling up, high expression level, proper protein folding, fast growth rate and low cost, which makes them widely used in industry and research field.
Saccharomyces cerevisiae and the methylotrophic yeast Pichia pastoris are commonly used expression strain. Pichia pastoris is more popular, especially in the large-scale production of eukaryotic recombinant proteins, due to the fast growth rate, multiple commercial expression vectors and high efficiency of secretion expression.
Royobiotech Co.,Ltd provides a well-developed yeast expression and purification services, including the mini-scale and large scale expression and purification of recombinant protein in Pichia pastoris and Saccharomyces cerevisiae.
Service:
Recombinant Plasmid Generation for Yeast Expression
Transformation of Linear Construct
Verification of Positive Transformants by PCR and Western Blot Method
Small Scale Expression and Purification (2L cell culture)
Scale Up Expression and Purification of Recombinant Protein (10L or more)
|
Steps |
Items |
Timeline |
|
1 |
Recombinant Plasmid Generation |
2 weeks |
|
2 |
Transformation of Linear Construct |
1 week |
|
3 |
Verification of Positive Transformants via PCR |
3-4weeks |
|
4 |
Small Scale Expression and Purification |
2-4 weeks |
|
5 |
Scale Up Expression and Purification |
TBD |
Remarks:
The service is highly flexible to meet your request. One or more above steps could be separately provided.
Affinity purification, ion exchange purification, hydrophobic beads purification and size exclusion column purification or their combination will be used for protein purification.
Baculovirus Expression Vector System
Baculovirus expression vector system (BEVS) is a power system to produce a variety of prokaryotic and eukaryotic protein. It is also an efficient tool for preparing vaccines.
The system could effectively produce high yield target protein with proper folding and post-translation modifications in an intracellular or secretion expression means.
Royobiotech Co.,Ltd researchers establish a well-developed baculovirus-insect cell expression services platform. We provide the expression and purification services on preparation of recombinant baculovirus and recombinant protein. We have a good track record in producing kinase and recombinant protein complexes.
Service：
Generation of recombinant Bacmid DNA
Preparation of recombinant baculovirus
Titration of baculovirus
Protein expression verification and optimization
Small scale expression and purification of recombinant protein in insect cell
Scale up expression and purification of recombinant protein in insect cell
|
Steps |
Items |
Timeline |
Comments |
|
1 |
Generation of recombinant Bacmid DNA |
3 weeks |
Different affinity tag: GST tag, His tag, Flag tag….. |
|
2 |
Preparation of recombinant baculovirus |
2 weeks |
|
|
3 |
Titration of baculovirus |
2-3 days |
Quantitative-PCR or based on gp64 antibody |
|
4 |
Protein expression verification and optimization |
2 weeks |
MOIs, PIs or different cell lines |
|
5 |
Small Scale expression and purification |
2-4 weeks |
500 ml-2L expression in flask |
|
6 |
Scale up expression and purification |
TBD |
Culture in flask or fermentor |
Remarks:The service is highly flexible to meet your request. One or more above steps could be separately provided.
Mammalian Expression
CHO, Vero, BHK, 293 cells etc
Adherence and Suspension Culture
Transient Expression Screen
Stable Cell Line Generation
Rabbit IgG expressed in transient transfected 293 cells
