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Recombinant Protein Expression and Purification Services

2016-10-12
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Medicilon offers Recombinant Protein Expression and Purification Services. Medicilon is a leading provider of comprehensive, high quality recombinant protein and bioprocess services. We offer a variety of recombinant protein expression platforms along with a host of other protein services like chemical protein synthesis, protein refolding and structural biology services.

Medicilon offers extensive expertise across four expression systems for producing your recombinant proteins, monoclonal antibodies, vaccine antigens or disease biomarkers. Whether for structural studies, functional assays, target validation or high throughput screens, we can help.

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Protein Expression System

  • Mammalian Expression

Applications Requiring Protein Expression and Purification

  • Enzymology / Functional Studies (mg)

  • Antibody Production (ug-mg)

  • Structure Determination

  • Commercial / Therapeutic Uses (g)

Types of Protein to be Expressed and Purified

  • Soluble: Cytoplasmic; Easiest to work with; Usually no Soluble

  • Secreted (stable) : Proteases, Antibodies, Cytokines; Frequently contain PTMs

  • Membrane Associated: Intracell or Extracell

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.

 

Medicilon researchers established a well-developed E. coli expression and purification services platform, which offers the expression and purification services of a variety of recombinant proteins, with a fast turn-around time and competitive pricing.

 

Services:

  1. Generation of Recombinant Plasmid

  2. Expression Test in Host

  3. Soluble Expression Test

  4. Small Scale Expression and Purification (2L Cell Culture)

  5. Scale Up Expression and Purification

StepsItemsTimelineComments
1Generation of Recombinant Plasmid2 weeksMultiple expression vector, different affinity tag: His, Trx , GST, SUMO…
2Expression of Recombinant Protein1 weekMultiple expression host
3Soluble Expression Optimization1-2 weeksTest different induction concentration, time point, temperatures or reconstruct plasmids
4Small Scale Expression and Purification1-2 weeksPurification of soluble expression or from inclusion body
5Scale Up Expression and PurificationTBDCulture in flask or fermenter

Remarks:

  1. The service is highly flexible to meet your request. One or more above steps could be separately provided.

  2. 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.

 

Medicilon 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.

 

Services:

  1. Recombinant Plasmid Generation for Yeast Expression

  2. Transformation of Linear Construct

  3. Verification of Positive Transformants by PCR and Western Blot Method

  4. Small Scale Expression and Purification (2L cell culture)

  5. Scale Up Expression and Purification of Recombinant Protein (10L or more)

StepsItemsTimeline
1Recombinant Plasmid Generation2 weeks
2Transformation of Linear Construct1 week
3Verification of Positive Transformants via PCR3-4weeks
4Small Scale Expression and Purification2-4 weeks
5Scale Up Expression and PurificationTBD

Remarks:

  1. The service is highly flexible to meet your request. One or more above steps could be separately provided.

  2. 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.

 

Medicilon 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.

 

Services:

  1. Generation of recombinant Bacmid DNA

  2. Preparation of recombinant baculovirus

  3. Titration of baculovirus

  4. Protein expression verification and optimization

  5. Small scale expression and purification of recombinant protein in insect cell

  6. Scale up expression and purification of recombinant protein in insect cell

StepsItemsTimelineComments
1Generation of recombinant Bacmid DNA3 weeksDifferent affinity tag: GST tag, His tag, Flag tag…..
2Preparation of recombinant baculovirus2 weeks 
3Titration of baculovirus2-3 daysQuantitative-PCR or based on gp64 antibody
4Protein expression verification and optimization2 weeksMOIs, PIs or different cell lines
5Small Scale expression and purification2-4 weeks500 ml-2L expression in flask
6Scale up expression and purificationTBDCulture 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

 

Rabbit IgG expressed in transient transfected 293 cells

Rabbit IgG expressed

Note: The condition medium was run in 10%SDS-PAGE gel and stained with coomassie blue

 

 

Contact Us 

Email : marketing@medicilon.com

Tel : +86 021 58591500

 

Tips : Above is part of recombinant protein purification, Protein Expression and Purification. You can also CONTACT US with any question or enquiry you may have. We will be happy to discuss your needs in detail and design an appropriate plan of action.

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