Fractionation of bovine serum albumin and monoclonal antibody alemtuzumab using carrier phase ultrafiltration.
Protein transmission and hence selectivity of separation can be significantly affected by solution pH and ionic strength in protein fractionation using ultrafiltration. Using parameter scanning ultrafiltration, the transmission of bovine serum albumin (BSA) and monoclonal antibody alemtuzumab (Campath-1H) through 300 kDa polyethersulfone (PES) ultrafiltration membranes were studied over a range of pH and salt concentrations, with focus on the likely conditions for achieving "reverse selectivity," i.e., obtaining purified alemtuzumab (approximately 155 kDa) in the permeate. Experimental results demonstrate that the reverse selectivity could be obtained by manipulating the operating conditions such as the solution pH, ionic strength, permeate flux, and system hydrodynamics. With a two-stage batch ultrafiltration process under suitable conditions, the monoclonal antibody alemtuzumab with a purity of > 98% was obtained in the permeate from a feed solution initially containing 0.50 g/l each of BSA and alemtuzumab. Further purity can be expected by selecting more suitable membranes and optimizing operating conditions.
Wan
Yinhua
Y
Department of Engineering Science, University of Oxford, Oxford OX1, 3PJ, United Kingdom.
Ghosh
Raja
R
Hale
Geoff
G
Cui
Zhanfeng
Z
D000074323:Alemtuzumab; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000302:isolation & purification*; D061067:Antibodies, Monoclonal, Humanized; D000912:Antibodies, Neoplasm / Q000737:chemistry* / Q000302:isolation & purification*; D005591:Chemical Fractionation / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D008567:Membranes, Artificial; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000302:isolation & purification*; D012965:Sodium Chloride / Q000737:chemistry; D014462:Ultrafiltration / Q000379:methods*
D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D000912:Antibodies, Neoplasm; D008567:Membranes, Artificial; D012710:Serum Albumin, Bovine; D000074323:Alemtuzumab; D012965:Sodium Chloride