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Case Studies

Case Studies

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)
OptimumGene™ Codon Optimization Technology     See more  
Co-expression of Protein Complexes     See more  
BacPower™ Bacterial Expression System     See more  
YeastHIGH™ Yeast Expression System      See more  
BacuVance™ Baculovirus Expression System     See more  
Mammalian Expression System     See more  

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