Pichia Protein Expression Validation (LabChip)

URL: https://cloud.ginkgo.bio/protocols/pichia-protein-expression-labchip Status: Ginkgo Certified (Newly launched) Price: $89/sample Turnaround: up to 4 weeks Throughput: Up to 96 constructs per run

Overview

End-to-end automated workflow using Ginkgo's proprietary Pichia pastoris strains to express and characterize secreted proteins. Strains progress through three sequential preculture and production fermentation stages (multi-stage fed-batch-style growth). Secreted protein expression is quantified by size and purity via microfluidic capillary electrophoresis (LabChip). Useful for targets that express better in a eukaryotic secretion system than in E. coli or cell-free.

Input

• DNA Input: Linear DNA sequence (.xlsx template)

Output

• Expression Confirmation: Verification of the target protein at the expected molecular weight
• Baseline Titer: Initial quantitative yield measurement (mg/L)
• Initial Purity: Percentage of target protein vs. impurities, delivered with virtual gel images
• Results reported as protein size (kDa), purity (%), and relative concentration

Automated Workflow

1. Transformation using 384-well plates
2. Multi-stage fermentation (three sequential preculture and production stages, fed-batch-style growth)
3. Supernatant harvest by high-speed centrifugation
4. Quantification Prep - dispense PBS diluent (BioTek MultiFlo), seal (Agilent PlateLoc), store at 4 deg C (HRB TundraStore)
5. LabChip protein quantification (size, purity, relative concentration)

Ordering

• Number of Proteins: configurable
• Number of Replicates: configurable
• File Upload: CSV, Excel, FASTA, TXT, PDF, ZIP
• Additional Details: free-text field for special requirements

Use Cases

• Expressing secreted/eukaryotic proteins that are difficult in E. coli or cell-free
• Validating expressibility in Pichia pastoris before scale-up
• Size/purity profiling of secreted targets