Parameter scanning ultrafiltration: rapid optimisation of protein separation.
High-resolution fractionation of proteins using ultrafiltration is feasible only at highly optimised conditions. Conventional process optimisation methodology demands both time and material. Pulsed sample injection ultrafiltration has been suggested as a rapid process optimisation technique. In the present work the scope of this technique is further extended by "parameter scanning ultrafiltration," which involves continuous change of a process parameter (e.g., pH, salt concentration). The time and material consumption are thus further reduced. The technique was validated using different proteins and membranes. Sieving coefficients at different pH and salt concentration were compared to those obtained in fixed parameter ultrafiltration experiments. As fractionation case studies the separation of monoclonal antibody from bovine serum albumin and separation of human IgG from human serum albumin were examined.
Ghosh
Raja
R
Department of Chemical Engineering, McMaster University, 1280 Main Street West, Hamilton, Ontario, Canada. rghosh@mcmaster.ca
Wan
Yinhua
Y
Cui
Zhanfeng
Z
Hale
Geoff
G
D000074323:Alemtuzumab; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification; D061067:Antibodies, Monoclonal, Humanized; D000912:Antibodies, Neoplasm / Q000737:chemistry / Q000302:isolation & purification; D002417:Cattle; D005591:Chemical Fractionation / Q000295:instrumentation / Q000379:methods*; D002846:Chromatography, Affinity / Q000379:methods; D004867:Equipment Design; D005246:Feedback; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000737:chemistry / Q000302:isolation & purification; D009113:Muramidase / Q000737:chemistry / Q000302:isolation & purification; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D011786:Quality Control; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012709:Serum Albumin / Q000737:chemistry / Q000302:isolation & purification; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000302:isolation & purification; D014462:Ultrafiltration / Q000295:instrumentation / Q000379:methods*
D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D000912:Antibodies, Neoplasm; D007074:Immunoglobulin G; D011506:Proteins; D012709:Serum Albumin; D012710:Serum Albumin, Bovine; D000074323:Alemtuzumab; D009113:Muramidase