Elution of proteins from hydroxyapatite resins without resin deterioration
AssigneeBio-Rad Laboratories, Inc.
InventorLarry J. Cummings
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Abstract
Proteins, including monoclonal antibodies, that have been retained on hydroxyapatite resins for purposes of protein separation, purification, or both, are eluted from the resins by a elution buffer that contains controlled amounts of calcium and phosphate ions. The buffer allows elution to be performed in repeated runs at an acidic pH without deterioration of the resin.
Claims11
A method for eluting proteins from a solid phase comprising hydroxyapatite to which said proteins are bound, said method comprising passing through said solid phase an eluent comprising calcium ion at a concentration of from about 25 ppm to about 260 ppm and phosphate ion at a concentration of from about 2 mM to about 40 mM at a pH of from about 5.3 to about 5.8.
A method for extracting monoclonal antibodies from high-molecular-weight aggregates in a solution comprising said antibodies and aggregates, said method comprising:
- (i) applying said solution to a solid phase comprising hydroxyapatite, and
- (ii) passing through said solid phase an eluent comprising calcium ion at a concentration of from about 50 ppm to about 225 ppm, phosphate ion at a concentration of from about 5 mM to about 40 mM, and an alkali metal salt at a concentration of from about 0.3M to about 1.5N, at a pH of from about 5.3 to about 5.8 to obtain an eluate containing said monoclonal antibodies.
Dependencies are inferred from the claim text.