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Recombinant Protein Expression in Yeast Cells

Similar to mammalian cells, yeast cells confer eukaryotic processing, folding, and post-translational modification. At the same time, a yeast expression system delivers high level expression. Bon Opus has established an efficient system for yeast cell recombinant protein expression, which includes the design of signal sequences, expression vectors, and a set of yeast strains (X33, GS115, KM71, and SMD1168). Our team incorporates the use of antibiotic gradients to screen for transformants with the highest numbers of inserts, and thus ensure a high expression level in the follow-up production phase.

Feature Package: TotumPro™ Yeast Protein Production ($2500 per 0.5mg)

  • Gene synthesis and cloning into yeast expression vector

  • Preliminary expression detection in yeast cells: 10 positive clones, SDS analysis, WB

  • 200ml fermentation & one-step purification (His) of optimal clone in yeast cells

  • 90% purity, tag-on protein, no endotoxin control

  • 6-8 weeks delivery time

General Workflow

1. Gene synthesis

2. Cloning & Transformation

3. Clone Selection & Expression

4. Purification

Steps Breakdown

Steps

Timeline

1) Gene synthesis, codon optimization, and DNA prep

2 weeks

2) Transformation into P. pastoris cells

1 week

3) Screening for transformants with multiple copies of inserts

1 week

4) Expression, evaluation, and optimization

2 weeks

5) Scale-up production

Inquiry

6) Final QC and delivery of product

Case Studies

Optimized Pichia pastoris Expression

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  • Strategy:

    • ​200 mL yeast culture expression

    • Purification using Ni-NTA column

    • QC by SDS-PAGE and anti-His WB

  • Deliverables: 

    • 6.6 mg protein 

    • Purity >95% by reducing SDS-PAGE

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Bon Opus Biosciences is an expert in recombinant protein expression with yeast host cells. We cater our comprehensive workflow to serve each application as it works for you. We guarantee accuracy and efficacy with high-achieving yields and robust production rates. We guarantee our yeast protein expression services function similarly to other host cells, so you can rest assured knowing your selection is accurate and handled properly. Our in-house lab experts and quality control team carry extensive industry education and knowledge and employs their expertise throughout each step of the process. When you’re in need of accurate, robust, and transparent workflows, look no further than Bon Opus Biosciences! 

The Purpose for Yeast Expression 
Yeast is an excellent host cell strain for recombinant eukaryotic expression demands because it’s fairly easy to manipulate during the workflow. Specific proteins are expressible with yeast because of this easy manipulation, and both heterologous and endogenous are included. With an easy workflow system, Bon Opus Biosciences can produce highly robust and production yeast host cell strain yields for a wide range of applications, including biopharmaceutical and industrial demands. Our yeast protein production workflow is distinct, with a hands-on approach for each application. 

Bon Opus Biosciences Distinct Workflow Design 
Our team of in-house lab and quality control experts work diligently to complete a thorough six-step process. In each step, we take you along with us to ensure your satisfaction and resolution should any issues arise. To begin our custom recombinant protein production in yeast cells, we spend two weeks synthesizing the cells, undergoing codon optimization, and preparing the DNA. We house a distinct inventory of preclinical DNA prep that can work for your application. After thorough completion of the synthesizing, we spend one week transforming the genes to P. pastoris cells. This allows us to spend time screening the production for copies. From here, we spend two weeks expressing, evaluating, and optimizing. Optimization is imperative in most custom services because it allows us to use fusion tags that offer custom and robust yields. We will take the fully optimized protein and scale-up with purification as it suits the protein of interest. At the end of all yeast protein production services, we complete our in-house quality control and prepare the yields for on-time delivery. 

 

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