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Protein Expression and Purification


IN BACTERIA IN BACTERIA
   
IN INSECT CELLS (BACULOVIRUS) IN INSECT CELLS (BACULOVIRUS)
   
IN MAMMALIAN CELLS IN MAMMALIAN CELLS

Options for Protein Expression Services


ProAlt offers protein expression services from gene cloning to large-scale production to speed up your research. Our experienced staff ensures you the highest standards in protein expression and purification. Our services are custom-tailored to meet the needs of industry, academic and government researchers.


The choice of a suitable service depends on many factors. These include your requirements in relation to economical cost, speed of your projects, amounts of the purified protein and mainly the post-translational modification profiles of your proteins and its applications.


Expression System Advantages Disadvantages
Bacteria
  • Low cost
  • Fast
  • Easy manipulation and scale up
  • Possible misfolding of proteins
  • Lack of post-translational modifications
  • Different codon usage
Baculovirus
  • Eukaryotic environment
  • High levels of recombinant gene expression
  • Easy of scale-up
  • Discontinuous expression
  • Less rapid and more costly than prokaryotic
Mammalian
  • Highest level of protein with post-translational processing
  • Low protein yield
  • Less rapid and more expensive than bacteria and baculovirus



En bacterias PROTEIN EXPRESSION IN BACTERIA

Modularized Service: all the steps from cloning to protein purification are available independently to meet your specialized requirements.


Bacterial expression service request


Reference Project Phase / Description Estimated time
ECO-01 Ia. Cloning into bacterial expression vector (1 tag) 2 weeks
ECO-02 Ib. Cloning into 2 bacterial expression vectors (2 different tags)
ECO-03 Ic. Cloning into 3 bacterial expression vectors (3 different tags)
ECO-04 IIa. Small scale expression test (1 tag) 1-2 weeks
ECO-05 IIb. Small scale expression test (2 different tags)
ECO-06 IIc. Small scale expression test (3 different tags)
MALDI MALDI-TOF: Peptide mass fingerprinting 1 week
ECO-07 III. Large scale production (1 to 5 liters of bacterial culture) 3-5 days
ECO-08 IVa. Purification of recombinant soluble protein 1 week
ECO-09 IVb. Purification of insoluble His-tagged protein

Tags: 6xHis; GST; MBP; others


NOTES:


  • Customer will provide to ProAlt the DNA/gen containing the whole nucleotide sequence (100%) of the protein to be expressed (sequence verified; without introns, mutations, deletions and/or internal stop codons).
  • Customer can additionally request to ProAlt the gen synthesis service, including codon optimization for protein expression in a selected system (bacterial, baculovirus, mammalian, others). For this purpose, customer will provide to ProAlt the whole nucleotide and/or aminoacid sequence for the protein of interest and will receive a separately quote related to this additional service.
  • As a proper expression depends on the protein itself, a successful production and/or the yield or purity of each batch of the protein of interest can not be guaranteed in advance. Therefore, you will only be invoiced for the Project Phase completed. Large-scale culture may be needed to obtain a certain amount of protein for antibody production.
  • Estimated times are given as a guide and will suffer modifications according to complexity and stability of each protein during design and production.
  • Modules ECO-08 or ECO-09 can not be selected in advance since the purification method of the recombinant protein of interest (soluble or insoluble) depends on the protein nature and properties. This method will be determined previously in module ECO-04.

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PROTEIN EXPRESSION IN INSECT CELLS (BACULOVIRUS) PROTEIN EXPRESSION IN INSECT CELLS (BACULOVIRUS)

Modularized Service: all the steps from cloning to protein purification are available independently to meet your specialized requirements.


Baculovirus expression service request


Reference Project Phase / Description Estimated time
BCV-01 Ia. Cloning into baculoviral expression vector (1 tag) 2 weeks
BCV-02 Ib. Cloning into two baculoviral expression vectors (2 different tags)
BCV-03 IIa. Generation of recombinant baculovirus (1 tag) 2-3 weeks
BCV-04 IIb. Generation of recombinant baculovirus (2 different tags)  
MALDI MALDI-TOF: Peptide mass fingerprinting 1 week
BCV-05 III. Amplification of recombinant baculovirus for high titer stock 1-2 weeks
BCV-06 IV. Expression optimization 1-2 weeks
BCV-07 V. Large scale production (500 ml to 5 liters of Sf9 insect cells) 1 week
BCV-08 VIa. Purification of recombinant soluble protein 1 week
BCV-09 VIb. Purification of insoluble His-tagged protein

Tags: 6xHis and/or GST


NOTES:


  • Customer will provide to ProAlt the DNA/gen containing the whole nucleotide sequence (100%) of the protein to be expressed (sequence verified; without introns, mutations, deletions and/or internal stop codons).
  • Customer can additionally request to ProAlt the gen synthesis service, including codon optimization for protein expression in a selected system (bacterial, baculovirus, mammalian, others). For this purpose, customer will provide to ProAlt the whole nucleotide and/or aminoacid sequence for the protein of interest and will receive a separately quote related to this additional service.
  • As a proper expression depends on the protein itself, a successful production and/or the yield or purity of each batch of the protein of interest can not be guaranteed in advance. Therefore, you will only be invoiced for the Project Phase completed. Large-scale culture may be needed to obtain a certain amount of protein for antibody production
  • Estimated times are given as a guide and will suffer modifications according to complexity and stability of each protein during design and production.
  • Modules BCV-08 or BCV-09 can not be selected in advance since the purification method of the recombinant protein of interest (soluble or insoluble) depends on the protein nature and properties. This method will be determined previously in module BCV-06.

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PROTEIN EXPRESSION IN MAMMALIAN CELLS PROTEIN EXPRESSION IN MAMMALIAN CELLS

Modularized Service: all the steps from cloning to protein purification are available independently to meet your specialized requirements.


Mammalian expression service request


Reference Project Phase / Description Estimated time
MAM-01 I. Cloning into mammalian expression vector 2 weeks
MAM-02 II. Small scale expression test 2 weeks
MALDI Espectrometría de masas MALDI-TOF 1 semana
MAM-03 III. Medium scale production (108 mammalian cells) 2 weeks
MAM-04 IVa. Purification of recombinant soluble protein 2 weeks
MAM-05 IIVb. Purification of insoluble His-tagged protein 1 semana

Tags: 6xHis


Cells: 293-EBNA y CHO-K1 (adherent); 293 y CHO (suspension)


NOTES:


  • Customer will provide to ProAlt the DNA/gen containing the whole nucleotide sequence (100%) of the protein to be expressed (sequence verified; without introns, mutations, deletions and/or internal stop codons).
  • Customer can additionally request to ProAlt the gen synthesis service, including codon optimization for protein expression in a selected system (bacterial, baculovirus, mammalian, others). For this purpose, customer will provide to ProAlt the whole nucleotide and/or aminoacid sequence for the protein of interest and will receive a separately quote related to this additional service.
  • As a proper expression depends on the protein itself, a successful production and/or the yield or purity of each batch of the protein of interest can not be guaranteed in advance. Therefore, you will only be invoiced for the Project Phase completed. Large-scale culture may be needed to obtain a certain amount of protein for antibody production
  • Estimated times are given as a guide and will suffer modifications according to complexity and stability of each protein during design and production.
  • Modules MAM-04 or MAM-05 can not be selected in advance since the purification method of the recombinant protein of interest (soluble or insoluble) depends on the protein nature and properties. This method will be determined previously in module MAM-02.

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