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研究例


Protein with Biological Activity

Production and purification of proteins with biological activity is essential in drug discovery. Multifunctional units at GenScript secure successful delivery of bioactive proteins based on our state-of-the-art equipment and methodologies. A case study below shows groups of protein production and assay development teaming up deliver enzyme with qualified specific activity, higher than customer expected. After modifying the method of cell-breaking, we increase the final yield of protein of interest with purity of above 90%.

Biological Activity Proteins
Fig. A: SDS-PAGE analysis of the fractions upon the IEX chromatography
Lane 1-47: Gradient elution fractions
Lane M: Smart Broad-Range Protein Standard (GenScript, Cat. No. MM0906)
Lane S: The supernatant after homogenizer with high pressure
Lane ft: Flow through
Biological Activity Proteins
Fig. B: Enzyme assay monitoring the fractionation by DEAE Sepharose
Biological Activity Proteins
Fig. C: Fractions pooled based on activity analysis for further ADP Sepharose affinity purification
Fig. D: SDS-PAGE analysis of the purified protein

Lane M: Smart Broad-Range Protein Standard (GenScript, Cat. No. MM0906)
Lane S: The supernatant after homogenizer with high pressure
Lane ft: Flow through

Large-scale Endotoxin Free Protein Production

GenScript's protein team has extensive experience in bacterial expression and purification, protein refolding, solubility enhancement, endotoxin removal, and large-scale production.

In a recent case, we have obtained 108 mg A fusion protein with >90% purity from 20 L E. coli expression and two-step purification. The concentration is 4.02 mg/ml and the endotoxin level is lower than 0.8 EU/ug.

  Fig. 1 SDS-PAGE analysis of the expression of A fusion protein   Fig. 2 SDS-PAGE analysis of the first step purification of A fusion protein  
  Lane M: Smart Broad-Range Protein Standard (Genscript, Cat.No. MM0906)   Lane 1: Supernatant fraction Lane 3: Wash  
Lane 2: Flow through fraction Lane 4–5: Elution  
  Lane 1: Lysate of cells without induced   Lane M: Smart Broad-Range Protein Standard (Genscript, Cat.No. MM0906)  
  Lane 2–4: Lysate of cells induced  
  Fig.3 SDS-PAGE analysis of the second step purification of A fusion protein   Fig. 4 SDS-PAGE analysis of the purity of A fusion protein  
Lane 1: A fusion protein
Lane M: Smart Broad-Range Protein Standard (Genscript, Cat.No. MM0906)
  Lane 1: Specimen fraction Lane 2–9: Elution    
Lane M: Smart Broad-Range Protein Standard (Genscript, Cat.No. MM0906)

   
    Co-expression of Protein Complexes

Purification of macromolecular complex subunits in large quantities and assembling them into functional entities are vital steps for successfully determining the structure and characterizing the functions of macromolecular complexes. Despite the fact that cases of isolation and structure determination of endogenous complexes have been demonstrated, great efforts still lie ahead to make this technology easily accessible. Recent endeavor on this subject suggests that co-expression of subunits within hosts of E.coli and insect cells, even for high-throughput projects, is becoming a promising solution.

GenScript has successfully co-expressed 3 subunits of A protein in E. coli system:

Co-expression of protein complexes

Fig. A protein with His-tag was analyzed by SDS-PAGE followed by Coomassie Brilliant Blue staining
Lane M: Smart Broad-Range Protein Standard (GenScript, Cat. No. MM0906)
Lane 1: A-His-αβγ target protein with His-tag


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recombinant protein expression quotation

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recombinant protein expression To order, please download and complete the Quotation/Order Form and send it to us by email or fax with a formal PO (Purchase Order). You can also submit PO information by phone.

recombinant protein expression If submitting samples, please mail them together with a hard copy of the completed Quotation/Order Form to GenScript Japanese brach.

recombinant protein expression For questions about recombinant protein expression, or to inquire about the status of your order contact us by email, phone or fax.

Email: japanmarket@genscript.com
Phone: 03-6803-0843
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