Expression of proteins in yeast is a common alternative to prokaryotic and higher eukaryotic expression. Yeast cells offer many of the advantages of producing proteins in microbes (growth speed, easy genetic manipulation, low cost media) while offering some of the attributes of higher eukaryotic systems (post translational modifications, secretory expression). KMD Bioscience offers comprehensive, high quality recombinant custom protein services from yeast expression systems. Our goal is to provide a flexible service program tailored to your needs every step of the way. We offer a complete ‘start to finish’ service from the cDNA (gene) cloning step to the protein purification step, and downstream modifications such as tag addition-removal, endotoxin testing/removal, conjugations, and any other services that you may request.
Yeast Protein Expression Service Procedure:
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Steps
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Specification
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Timeline
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Deliverables
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Gene synthesis, Cloning, Plasmid Prep
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* Gene synthesis and Codon optimization
* Sub cloning into appropriate expression vector
* Plasmid Prep and DNA QC
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1 week
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* Recombinant expression plasmid
* DH5α strains
* Sequencing report
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Transformation of yeast expression strains
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* Transform plasmids into appropriate yeast expression strains
* Amplification and culture of the above 5-10 positive clones
* SDS-PAGE
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2 weeks
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* Positive clone strains
* Screening report
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Screening of yeast expression strains
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* Detection of recombinant protein expression
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2 weeks
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* Positive clone strains
* Screening report
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Protein expression evaluation and optimization
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* Scale up protein expression using optimized conditions
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2 weeks
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Small-scale protein expression and purification
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* The recombinant proteins were purified by affinity chromatography, ion exchange chromatography and gel filtration (1L yeast solution)
* SDS-PAGE
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1-2 weeks
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* 1~10mg recombinant protein
* Protein purification report
* Tag removal and separation of tag-free protein (if requested)
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Large-scale protein expression and purification
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* 5L yeast solution（According to client requirements）
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1-2 weeks
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* According to client's requirements
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Value-added services
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* Detection of the quality of the target protein by HPLC, mass spectrometry（MS）
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1-2 weeks
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* According to client's requirements
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Service Highlights:
*Codon optimization: including rare codon analysis, protein hydrophilicity analysis and prediction of transmembrane domains.
*A variety of fusion tag vectors and hosts to ensure high soluble protein expression and activity.
*Scale-up: from 500ml to 500L.
*Short cycle and high output.
*LAL <0.1EU/mg.
*Customization: quantity, purity, tags, and endotoxin levels are customizable.
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