Hydroxyapatite chromatography: purification strategies for recombinant proteins. 2014

Larry J Cummings
Bio-Rad Laboratories, Inc., Hercules, CA, USA. Electronic address: larry_cummings@bio-rad.com.

The purpose of this section is to provide developmental purification strategies for recombinant proteins on a column of hydroxyapatite and then to optimize them for repeated purification cycles using effect additives. Surface neutralization solutions (SNS) (patent pending) and/or the addition of calcium ions reduce the solubility of hydroxyapatite thus extending the column lifetime. This section leads the user through purification protocol screening followed by SNS--calcium ion supplementation step gradients for purifying a monoclonal antibody from its contaminants. The section provides simple analytical tools using a 160 ml scale column to predict column life at the process scale. However, the development engineer is encouraged to test the model in a minimum-sized process column.

UI MeSH Term Description Entries
D011994 Recombinant Proteins Proteins prepared by recombinant DNA technology. Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D002118 Calcium A basic element found in nearly all tissues. It is a member of the alkaline earth family of metals with the atomic symbol Ca, atomic number 20, and atomic weight 40. Calcium is the most abundant mineral in the body and combines with phosphorus to form calcium phosphate in the bones and teeth. It is essential for the normal functioning of nerves and muscles and plays a role in blood coagulation (as factor IV) and in many enzymatic processes. Coagulation Factor IV,Factor IV,Blood Coagulation Factor IV,Calcium-40,Calcium 40,Factor IV, Coagulation
D002853 Chromatography, Liquid Chromatographic techniques in which the mobile phase is a liquid. Liquid Chromatography
D004867 Equipment Design Methods and patterns of fabricating machines and related hardware. Design, Equipment,Device Design,Medical Device Design,Design, Medical Device,Designs, Medical Device,Device Design, Medical,Device Designs, Medical,Medical Device Designs,Design, Device,Designs, Device,Designs, Equipment,Device Designs,Equipment Designs
D006863 Hydrogen-Ion Concentration The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH pH,Concentration, Hydrogen-Ion,Concentrations, Hydrogen-Ion,Hydrogen Ion Concentration,Hydrogen-Ion Concentrations
D000911 Antibodies, Monoclonal Antibodies produced by a single clone of cells. Monoclonal Antibodies,Monoclonal Antibody,Antibody, Monoclonal
D017886 Durapatite The mineral component of bones and teeth; it has been used therapeutically as a prosthetic aid and in the prevention and treatment of osteoporosis. Calcium Hydroxyapatite,Hydroxyapatite,Hydroxylapatite,Alveograf,Calcitite,Interpore-200,Interpore-500,Osprovit,Ossein-Hydroxyapatite Compound,Ossopan,Osteogen,Periograf,Hydroxyapatite, Calcium,Interpore 200,Interpore 500,Interpore200,Interpore500,Ossein Hydroxyapatite Compound

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