Fully Automated Sample Processing and HTP ... - PerkinElmer
Fully Automated Sample Processing and HTP ... - PerkinElmer Fully Automated Sample Processing and HTP ... - PerkinElmer
HTP Growth and Expression 2 plasmids 4+ promoters 3 ribosome binding sites 25 secretion leaders • High throughput parallel processing • Yield, quality and speed create significant advantages in real and opportunity costs 30 chaperone/disulfide bond isomerase overexpression plasmids 100+ protease clean deletion mutants, multiple deletions Strain Construction + plasmids host strains Seed electroporate and grow transformants on selective media HTP Expression Inoculate seed culture into HTP medium Replicate cultures will be grown for 24 h at 30 °C prior to induction Harvest and Analysis Thousands of strains evaluated in
Process analytical: 1 st and 2 nd tier screens challenge: to accurately analyze target in complex mixture of solids, protein, DNA, etc. in timely fashion Quantity addressed Quality addressed 1 st tier analysis 2 nd tier analysis Samples: 1000s Samples: 10s to 100s HTP Expression 30-50 OD Achieved HTP SDS-CGE HTP BLI MS HPLC 96-well format 90 min per plate 96-well format 30 min per plate + Western Blot and other methods measure target mass yield measure fragments, aggregates, etc. Discovery ● Development ● Production
- Page 1 and 2: Fully Automated Sample Processing a
- Page 3 and 4: Who we are: • Located in San Dieg
- Page 5 and 6: Pfēnex HTP Strain Development >85%
- Page 7 and 8: Periplasmic Targeting of Recombinan
- Page 9: Value of Protease Deficient Hosts P
- Page 13 and 14: Expression analysis: Sample Prepara
- Page 15 and 16: Robotically Enabled Sample Preparat
- Page 17 and 18: Structure of the influenza hemagglu
- Page 19 and 20: Strains Selected for Fermentation R
- Page 21 and 22: 4-mL Fermentation PhyTip-IMAC Solub
- Page 23 and 24: Downstream Processing Robotic Batch
- Page 25 and 26: Summary Extensive genomics capabili
- Page 27: Acknowledgements: Pfenex Lauren Alc
Process analytical: 1 st <strong>and</strong> 2 nd tier screens<br />
challenge: to accurately analyze target in complex mixture of solids, protein, DNA, etc. in timely fashion<br />
Quantity addressed<br />
Quality addressed<br />
1 st tier<br />
analysis<br />
2 nd tier<br />
analysis<br />
<strong>Sample</strong>s: 1000s<br />
<strong>Sample</strong>s: 10s to 100s<br />
<strong>HTP</strong> Expression<br />
30-50 OD Achieved<br />
<strong>HTP</strong> SDS-CGE<br />
<strong>HTP</strong> BLI<br />
MS<br />
HPLC<br />
96-well format<br />
90 min per plate<br />
96-well format<br />
30 min per plate<br />
+ Western Blot <strong>and</strong> other methods<br />
measure target mass yield<br />
measure fragments, aggregates, etc.<br />
Discovery ● Development ● Production
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