Caprylic acid-induced impurity precipitation from protein A capture column elution pool to enable a two-chromatography-step process for monoclonal antibody purification.
This article presents the use of caprylic acid (CA) to precipitate impurities from the protein A capture column elution pool for the purification of monoclonal antibodies (mAbs) with the objective of developing a two chromatography step antibody purification process. A CA-induced impurity precipitation in the protein A column elution pool was evaluated as an alternative method to polishing chromatography techniques for use in the purification of mAbs. Parameters including pH, CA concentrations, mixing time, mAb concentrations, buffer systems, and incubation temperatures were evaluated on their impacts on the impurity removal, high-molecular weight (HMW) formation and precipitation step yield. Both pH and CA concentration, but not mAb concentrations and buffer systems, are key parameters that can affect host-cell proteins (HCPs) clearance, HMW species, and yield. CA precipitation removes HCPs and some HMW species to the acceptable levels under the optimal conditions. The CA precipitation process is robust at 15-25°C. For all five mAbs tested in this study, the optimal CA concentration range is 0.5-1.0%, while the pH range is from 5.0 to 6.0. A purification process using two chromatography steps (protein A capture column and ion exchange polishing column) in combination with CA-based impurity precipitation step can be used as a robust downstream process for mAb molecules with a broad range of isoelectric points. Residual CA can be effectively removed by the subsequent polishing cation exchange chromatography.
Biologics Development Dept., Bristol-Myers Squibb, Bloomsbury, NJ, 08804.
Ji
J
Zheng
Biologics Development Dept., Bristol-Myers Squibb, Bloomsbury, NJ, 08804.
Lu
L
Wang
Biologics Development Dept., Bristol-Myers Squibb, Bloomsbury, NJ, 08804.
Barbara
B
Twarowska
Biologics Development Dept., Bristol-Myers Squibb, Bloomsbury, NJ, 08804.
Sarah
S
Laino
Biologics Development Dept., Bristol-Myers Squibb, Bloomsbury, NJ, 08804.
Colleen
C
Sparks
Biologics Development Dept., Bristol-Myers Squibb, Bloomsbury, NJ, 08804.
Timothy
T
Smith
Biologics Development Dept., Bristol-Myers Squibb, Bloomsbury, NJ, 08804.
Reb
R
Russell
Biologics Development Dept., Bristol-Myers Squibb, Bloomsbury, NJ, 08804.
Michelle
M
Wang
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification*; D016466:CHO Cells; D002210:Caprylates / Q000737:chemistry*; D011232:Chemical Precipitation; D002846:Chromatography, Affinity / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D006863:Hydrogen-Ion Concentration; D013696:Temperature
D000911:Antibodies, Monoclonal; D002210:Caprylates; C031492:octanoic acid
caprylic acid; impurity precipitation; monoclonal antibody; protein A capture column; two-chromatography step