Surface extenders and an optimal pore size promote high dynamic binding capacities of antibodies on cation exchange resins.
Increased recombinant protein expression yields and a large installed base of manufacturing facilities designed for smaller bulk sizes has led to the need for high capacity chromatographic resins. This work explores the impact of three pore sizes (with dextran distribution coefficients of 0.4, 0.53, and 0.64), dextran surface extender concentration (11-20mg/mL), and ligand density (77-138 micromol H+/mL resin) of cation exchange resins on the dynamic binding capacity of a therapeutic antibody. An intermediate optimal pore size was identified from three pore sizes examined. Increasing ligand density was shown to increase the critical ionic strength, while increasing dextran content increased dynamic binding capacity mainly at the optimal pore size and lower conductivities. Dynamic binding capacity as high as 200mg/mL was obtained at the optimum pore size and dextran content.
Hart
David S
DS
Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
Harinarayan
Chithkala
C
Malmquist
Gunnar
G
Axén
Andreas
A
Sharma
Mandakini
M
van Reis
Robert
R
D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification*; D002411:Cation Exchange Resins / Q000737:chemistry*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D003911:Dextrans / Q000737:chemistry; D009994:Osmolar Concentration; D010316:Particle Size; D016062:Porosity; D011485:Protein Binding
D000911:Antibodies, Monoclonal; D002411:Cation Exchange Resins; D003911:Dextrans