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Recombinant Insulin Optimization
Welcome
Progress is not how we grow; it is about how we make a difference in lives of others. This is our guiding principle at Yashraj Biotechnology Limited (YBL)
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Recombinant Insulin Optimization
System: E. coli
Scale: Lab (5 L) → Pilot (10 L) → Manufacturing (100 L)
Background
A client approached us with a recombinant insulin process expressed in E. coli. While inclusion bodies were formed efficiently, downstream recovery was inconsistent, and significant yield loss occurred during the refolding stage. The objective was to optimize induction, control inclusion body characteristics, and ensure reproducible scale-up to manufacturing volumes.
Key Challenges
- Variability in downstream recovery
- Yield loss during refolding
- Need for controlled inclusion body density rather than maximum expression
Our Approach
- Induction Strategy: High cell density induction (OD > 35) with reduced inducer concentration to balance expression and folding.
- Temperature Control: Shifted expression temperature to 30 °C, improving inclusion body consistency and reducing aggregation.
- Scale-Up Control: Applied oxygen transfer (kLa)–based scale-up rules, maintaining DO >35% across 5 L → 10 L → 100 L volumes.
Results
| Parameter | Outcome |
|---|---|
| Inclusion Body Consistency | High |
| Refolding Recovery | >70% |
| Yield Improvement | ~2.5–3× |
| Commercial Readiness | Achieved |
Impact
This optimization strategy enabled consistent downstream recovery, improved refolding efficiency, and delivered a commercially ready process. By focusing on inclusion body quality rather than sheer expression levels, the client achieved reliable manufacturing outcomes and reduced variability in product yield.
