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Yeast Expression System

Yeast protein expression service quotation

Yeast Expression System

GenScript's proprietary YeastHIGHTM technology combines the advantages of the bacterial and eukaryotic expression systems. Our yeast expression system employs stable and durable production strains that resemble mammalian protein processing, incorporating features of high yield, high productivity, and dramatic reductions in costs and time requirements.

GenScript has developed a platform that allows humanized antibodies to be produced in yeast expression system. The similarity between the yeast and mammalian system for recombinant antibody expression enables GenScript to reduce the high cost associated with manufacturing recombinant antibody in mammalian expression system.

In addition, as the largest gene synthesis supplier in US, GenScript can provide you with free advanced gene optimization specific to Pichia pastoris, Saccharomyces cerevisiae, and other yeast strains, to further enhance the productivity of your protein. Or use our free rare codon analysis tool to see if your gene sequence is suboptimal, and explore means to boost your target protein expression.

Grown in chemically defined media, the resulting purified secretory or intracellular proteins can be used for the following applications:

  • Eukaryotic protein analysis
  • NMR studies with isotope-labeled proteins
  • Assay screening
  • Structural biology studies
  • Animal feed production
  • Environmental engineering

  • Advanced Package
  • Standard Package
  • Antibody Production
  • Case Studies
  • GenScript Advantages
YeastHIGHTM Advanced Protein Expression Package (SC1346)

GenScript’s engineering-enhanced protein expression technologies enable a high success rate of over 90% over a wide variety of recombinant proteins, and our proprietary YeastHIGHTM technology is pushing yields even higher. Many secretory proteins that bacterial expression systems yield as insoluble or as inactive inclusion bodies can be produced successfully by our yeast expression system using native or yeast secretion signals.

Milestones Specifications Deliverables Timeline1
1. Subcloning
  • Subcloning into two appropriate expression vectors
  • Plasmid maxi preparation
  • Linearize the constructs
Construct(s) report
2 weeks
2. Expression optimization and evaluation
  • Transformation the linear construct into the appropriate host strain
  • Confirmation of positive transformants via PCR
  • Expression optimization and confirmation
    i: Screening of transformants with multiple gene-insert with in-house YeastFast technology
    ii: Media formulation and inducer concentrations
    iii: SDS-PAGE gel and/or Western blot
Expression data
3–5 weeks
3. Protein expression and purification
  • Small-scale expression from one liter
  • One-step purification, or additional purification to reach purity of 90% or higher
  • QC by SDS-PAGE, Western blot2, and MALDI-TOF3
Purified soluble protein
QC data
2 weeks
4. Large-scale protein production and purification (Optional)
  • Large-scale production up to 500 L (Employs fermentors and TFF large-scale filtration system)
  • Deliverables available at >90% purity or higher
  • Removal of tags
Yeast paste/
Purified proteins
QC data
2–3 weeks

1 Listed is the typical turnaround time at GenScript. Please note that additional requirements may incur additional steps, and likewise longer processing time.

2 FREE Western QC data is available for tagged proteins that can be detected by the anti-His or anti-GST antibody. For other tagged proteins, additional charges may apply.

3 Optional MALDI-TOF data is available with additional charge.

Standard Protein Expression Package (SC1257)

GenScript also provides a standard yeast expression package service, which can be conducted in accordance with any protocol the customer may see fit to supply.

Milestones Specifications Deliverables Timeline1
1. Subcloning
  • Subcloning into appropriate expression vector
  • Plasmid maxi preparation
  • Linearize the construct
Construct(s) report
2 weeks
2. Expression and evaluation
  • Transformation of the linear construct into the appropriate host strain
  • Confirmation of positive transformants via PCR
  • Expression confirmation
  • SDS-PAGE gel and/or Western blot
Expression data
2 weeks
3. Pilot expression and purification
  • 1 L yeast culture expression
  • One-step purification, or additional purification to reach purity of 90% or higher
  • QC by SDS-PAGE, Western blot2, and MALDI-TOF3
Purified proteins
QC data
2 weeks
4. Large-scale protein production and purification (Optional)
  • Large-scale production up to 500 L (Employs fermentors and TFF large-scale filtration system)
  • Deliverables available at >90% purity or higher
  • Removal of tags
Yeast paste/
Purified proteins
QC data
2–3 weeks

1 Listed is the typical turnaround time at GenScript. Please note that additional requirements may incur additional steps, and likewise longer processing time.

2 FREE Western QC data is available for tagged proteins that can be detected by the anti-His or anti-GST antibody. For other tagged proteins, additional charges may apply.

3 Optional MALDI-TOF data is available with additional charge.

Advantage of using yeast system for humanized antibodies production:

  • Animal free antibody production
  • More cost effective than mammalian expression system
  • Fast turnaround of large-scale antibody production

As in one case for demonstration, GenScript's scientists successfully expressed scFv, Fab, and full length Antibody A in yeast. Following figures reflect the expression results. All fragments were identified and confirmed by MALDI-TOF.

  Fig. B: SDS-PAGE analysis of anti-
A Fab expression

Band 1: Joint chain (24.5 kD + 23 kD);
Band 2: 24.5 kD; Band 3: 23 kD
 
Fig. A: SDS-PAGE analysis of anti-A Ig expression

Band 1: Heavy chain;
Band 2: Light chain

  Fig. C-1 (left figure): Western blot analysis of anti-A ScFv expression
Fig. C-2 (right figure): SDS-PAGE analysis of anti-A ScFv expression
1. Protein expression in yeast

GenScript's proprietary YeastHIGHTM expression system and in-house YeastFast technology delivers unparalleled advantages in secretory expression. Pictured below is an enzyme expression performed using our YeastHIGHTM expression technology. After one step purification by SP-Sepharose, we find tag-free proteins at an expression level of about 20 mg/L and at purity level of over 90%. The target protein is further identified by MALDI-TOF.

Yeast Expression System
Fig.     A: Expression and secretion of target protein from P.pastoris
Lane 1 : Conventinal yeast expression technology
Lane 2 : GenScript YeastHIGHTM technology
Lane 3 : Negative control
Fig.     B: Purification of the target protein by SP-Sepharose
Lane  M: Smart Protein Standard (GenScript, Cat. No. MM0900)

2. Signal peptides

Shown below is the expression of a protease with signal peptide. There was no secretion expression of target protein with Saccharomyces cerevisiae α factor prepro peptide. After changing the peptide sequence for secretion from α factor prepro to the native signal peptide of the protease itself, the target protein was expressed and secreted successfully.


Yeast Expression Signal Peptides
Fig.       A: Western blot analysis of target protein expression with native signal peptide
Lane 1-5: Induced for 24 h, 48 h, 72 h, 84 h, 120 h
Fig.       B: Western blot analysis of target protein expression with α factor prepro peptide
Lane 1-4: Induced for 24 h, 48 h, 72 h, 84h
Lane    M: Protein Marker
  1.YeastHIGHTM Technology
GenScript YeastHIGHTM technology harnesses the strongest advantages of yeast expression, including promoters, optimal copies of vector per cell, inducibility, signal peptides, and cellular location, allowing us to provide customers with high-level recombinant protein production. During our years in the yeast protein expression service field, we have successfully delivered hundreds of proteins, including enzymes, proteases, kinases, cytokines, receptors, and transcription factors, and among others.

2. OptimumGeneTM codon optimization technology for gene expression
GenScript's advanced OptimumGeneTM codon optimization technology can optimize your gene sequence, allowing it to reach the highest possible level of gene expression and protein solubility. The OptimumGeneTM algorithm optimizes a variety of parameters that are critical to transcription, translation, and protein folding. Learn more...

3. Large-scale protein production
GenScript's fermentor facility provides high-quality protein production, for orders of up to 500 L. Our TFF large-scale filtration system can enhance purity and yield.

4. Purification
GenScript offers a variety of flexible purification systems: affinity column, gel filtration, ion exchange, hydrophobic-interaction column, and HPLC. Various tags are added to protein to facilitate their purification and we also offer services for tag removal.

5. Competitive prices
Multiple yeast expression packages are supplied at competitive prices, starting from $2,900, to fit customer’s budget.

Quotations and Ordering:

protein service quotation

protein services To request a quotation, please download and complete the Quotation/Order Form, and send it to us by email or fax.

protein services 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) or credit card information. You can also submit PO/credit card information by phone or via our secure online messaging system.

protein services If submitting samples, please mail them together with a hard copy of the completed Quotation/Order Form to: Protein Services, GenScript, 120 Centennial Ave., Piscataway, NJ 08854, US.

protein services For questions or to inquire about the status of your order, contact us by email, phone, fax or via our secure online messaging system.

Our customer service representatives are available 24 hours a day, Monday through Friday to assist you.

Order by mail Email: protein@genscript.com
Order by phone Phone: 1-877-436-7274 (Toll-Free) 1-732-885-9188
Order by fax Fax: 1-732-210-0262 1-732-885-5878