By: Alan Schultz & Jack Bobzien.  Our company has developed a Fab fragment product and needed the purification process verified.  Using E. Coli, propose.

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Presentation transcript:

By: Alan Schultz & Jack Bobzien

 Our company has developed a Fab fragment product and needed the purification process verified.  Using E. Coli, propose a new purification train and determine cost evaluation.  Work within the given constraints.

 Affinity resins for Fab are too expensive for manufacturing scale.  Need to produce 50 kg/yr.  Expression levels are expected to reach 10% of total extracted protein in 10 years.  Achieve above a 99% purity in product.

 Fragment Antigen binding (Fab fragment)  Can be expressed in various hosts like E. Coli, yeast etc.  Periplasmic Expression  pI of 7.0  M.W. of 50 kDa

 E. Coli  Used Strain W3110  Widely used in the pharmaceutical industry.

 Immobilized metal-ion affinity chromatography  His 6 -tagged Fab  Use of EDTA-Mg 2+ and imidazole  92.4% purity in exit stream

 Overall: hrs  V-105: hrs  V-106: hrs

 Periplasmic Complications  For current system, an IMAC Column was major modification  $250 per g/ YFP