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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