1. By: Alan Schultz & Jack Bobzien

2.  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.

3.  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. https://www.venturacollege.edu/departments/academic/economics.shtml

4.  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 http://www.ruppweb.org/fab/fab_sketch_circled.gif

5.  E. Coli  Used Strain W3110  Widely used in the pharmaceutical industry. http://scm-l3.technorati.com/11/06/03/44263/e-coli-.jpg?t=20110603144325

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

15.  Overall: 64.74 hrs  V-105: 32.74 hrs  V-106: 26.21 hrs

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