Protein purification

CAFC
  • US 6,489,447 B1
  • Filed: 05/03/1999
  • Issued: 12/03/2002
  • Est. Priority Date: 05/06/1998
  • Status: Active Grant
First Claim
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1. A method for purifying a polypeptide from a composition comprising the polypeptide and a deamidated variant thereof, which method comprises the following steps performed sequentially:

  • (a) binding the polypeptide and deamidated variant to an ion exchange material using a loading buffer, wherein the loading buffer is at a first conductivity and pH;

    (b) washing the ion exchange material with an intermediate buffer at a second conductivity and/or pH so as to elute the deamidated variant from the ion exchange material;

    (c) washing the ion exchange material with a wash buffer which is at a third conductivity and/or pH, wherein the change in conductivity and/or pH from the intermediate buffer to the wash buffer is in an opposite direction to the change in conductivity and/or pH from the loading buffer to the intersediate buffer; and

    (d) washing the ion exchange material with an elution buffer at a fourth conductivity and/or pH so as to elute the polypeptide from the ion exchange material.

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