YeastHIGH^TM Yeast Expression System

Yeast protein expression system is the most economical eukaryotic expression system for both secretion and intracellular expression. It is ideally suitable for large-scale production of recombinant eukaryotic proteins.

Utilizing GenScript's YeastHIGH^TM Technology, selected stable and durable production strains that resemble mammalian system are employed for high-yield, high-productivity protein processing, thus dramatically reducing the associated costs and inherent time requirements. In addition, as the largest gene synthesis supplier in the U.S., GenScript can deploy free OptimumGene^TM Codon Optimization specific to Pichia pastoris, Saccharomyces cerevisiae, and other yeast strains to further enhance the productivity of your protein.

Combining advanced technologies and profound past experience, GenScript's YeastHIGH^TM Advanced Package enables high protein expression yield and achieves a success rate of over 90% for a variety of recombinant proteins. If you can provide protocol(s) for your protein expression and purification, YeastHIGH^TM Standard Package is recommended.

Advantages:

• Proprietary Technologies: Our YeastHIGH^TM Technology considers promoters, optimal copies of vector per cell, inducibility, signal peptides, and cellular location, which are essential for high-level recombinant protein production.

• Humanized antibody production: GenScript has developed a platform that produces humanized antibodies 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.

• Large-scale protein production: GenScript's fermentation facility provides high-quality protein production up to 2,000 L. The purity and yield can be greatly enhanced by our TFF large-scale filtration system.

• Streamlined Services: GenScript can take your project directly from gene synthesis, to downstream activities including recombinant protein expression, stable cell line generation, antibody production, and assay development.

• YeastHIGH^TM Advanced Package
• YeastHIGH^TM Standard Package
• Case Study
• Applications

YeastHIGHTM Advanced 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 2,000 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.

YeastHIGHTM Standard 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 2,000 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 Total price of YeastHIGHTM Standard Package: starting from $2,900 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.

Case Study—Humanized 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 View More Case Studies

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 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.

Quotations and Ordering:

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

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.

If submitting samples, please mail them together with a hard copy of the completed Quotation/Order Form to: Protein Services, GenScript, 860 Centennial Avenue., Piscataway, NJ 08854, US.

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.

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