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Service / Technology Instance

About Protein Production, Vestec near Prague, Czech Republic

View All Protein Production at Instruct

The core facility for protein production provides comprehensive services, covering every step from DNA to purified protein. These include gene cloning into expression vectors using various techniques and heterologous expression in Escherichia coli for bacterial expression, HEK293T, Expi293, and Expi293F™ GnTI- for mammalian expression, and the baculoviral system in Sf9 or High Five™ insect cells. The expression is followed by multistep protein purification.

 

Service Availability:

Remote

Physical

Instruments Available:

Cloning involves primer design and the preparation of DNA plasmids for protein expression using traditional restriction enzyme-based methods as well as restriction-free techniques. It also includes mutagenesis of existing plasmids. All constructs undergo verification by sequencing.

Literature review, cloning and vector design, protocol optimization.

We offer expression of recombinant proteins in several different expression systems.

Bacterial Expression in E. coli

We start with small-scale expression and solubilization tests to identify the optimal E. coli strain for your target protein. Based on these results, we move forward with large-scale expression cell harvesting.

Baculoviral Expression in Insect Cells

Using the baculoviral expression system in Sf9 or High Five™ insect cells, we begin with small-scale transfection and expression verification via western blot. Once confirmed, we amplify the baculovirus and scale up production for large quantities of your target protein.

Mammalian Expression Systems

We also employ mammalian expression systems using HEK293T/17SF, Expi293™, and Expi293F™ GnTI- cells. We first conduct small-scale transfection, verify expression by Western blot, and proceed to large-scale production.

We offer multistep purification for your target protein, utilizing a combination of affinity chromatography, ion exchange chromatography, and size exclusion chromatography. Our purification process ensures high purity and quality, and we deliver the protein at the desired concentration in aliquots. Along with the purified protein, we provide a detailed protocol of the whole procedure.

User Guide

  • Cloning services
    • Design of expression vectors
    • DNA template control by whole plasmid sequencing
    • Ordering gene fragments from an external company
    • Cloning using traditional restriction enzymes
    • Restriction-free methodologies (RF, KLD, In-Fusion)
    • Site-directed mutagenesis
    • Plasmid DNA preparation (minipreps, midipreps, maxipreps)
    • Preparation of competent cells
  • Recombinant protein production in E. coli
    • Small-scale expression and solubility tests in various E. coli strains
    • Large-scale protein expression
    • Protein production in minimal media
  • Recombinant protein production in insect cells
    • BacPAK™ (TaKaRa) and FlexiBAC baculovirus systems
    • Small-scale cell transfection and expression verification by Western blot
    • Baculovirus amplification
    • Large-scale production in Sf9 or High Five™ cells
  • Recombinant protein production in mammalian expression systems
    • Expression in HEK293T/17SF, Expi293™, Expi293F™ GnTI- cells
    • Small-scale cell transfection and expression verification by Western blot
    • Large-scale cell transfection and protein production
  • Recombinant protein purification
    • Affinity chromatography (HiTrap HP, Ni-INDIGO, Strep-Tactin®XT, GST, cobalt)
    • Ion-exchange chromatography
    • Size exclusion chromatography
    • Protein concentration and dialysis
    • Tag removal (TEV, SUMO, 3C protease)
  • Optimization of expression and purification protocols

Instruct Centre

BIOCEV

BIOCEV

Prumyslova 595

25250, Vestec, Prague

Czech Republic

www.biocev.eu

Protein Production, Vestec near Prague, Czech Republic

Contacts:

Lubica Skultetyova
Lubica Skultetyova
BIOCEV - Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University in Vestec
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Miroslava Alblova
Miroslava Alblova
Institute of Biotechnology
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