# US 10,501,491 B2 · Protein purification

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- Number: US 10,501,491 B2
- Document ID: US10501491B2
- Status: Granted
- Kind: B2
- Application number: 15/836,695
- Filed: 2017-12-08
- Published: 2018-08-30
- Granted: 2019-12-10
- Earliest claimed: 2002-09-11
- Assignees: Genentech, Inc.
- Applicants: Genentech, Inc.
- Inventors: Jefferson C. Emery; Paul J. McDonald; Rhona O'Leary
- Web page: https://patentgrep.com/patent/US10501491B2

## CPC

- C07K 1/18: Ion-exchange chromatography
- B01D 15/362: {Cation-exchange}
- B01D 15/363: {Anion-exchange}
- C07K 16/2863: {against receptors for growth factors, growth regulators}
- C07K 16/30: from tumour cells
- C07K 16/32: against translation products of oncogenes
- C07K 2317/21: from primates, e.g. man
- C07K 2317/565: Complementarity determining region [CDR]
- C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- B01D: SEPARATION (separating solids from solids by wet methods B03B, B03D; by pneumatic jigs or tables B03B; by other dry methods B07; magnetic or electrostatic separation of solid materials from solid materials or fluids, separation by high-voltage electric fields B03C; centrifuges, vortex apparatus B04; presses per se for squeezing-out liquid from liquid-containing material B30B9/02; treatment of water C02F, e.g. softening by ion-exchange C02F1/42; {arrangements of air intake cleaners in gas turbine plants F02C7/05} ; arrangements or mounting of filters in air-conditioning, air-humidification or ventilation F24F13/28)

## Abstract

A method for purifying a polypeptide by ion exchange chromatography is described in which a gradient wash is used to resolve a polypeptide of interest from one or more contaminants.

## Claims

### Claim 1

A method for purifying an antibody from a composition comprising the antibody and a contaminant, which method comprises the following steps performed sequentially:
(a) binding the antibody to a cation exchange material with an equilibration buffer at a first conductivity, wherein the amount of antibody in the composition loaded onto the cation exchange material is from about 15 mg to about 45 mg of the antibody per ml of cation exchange material;
(b) washing the cation exchange material with a wash buffer with a conductivity that increases at a first rate from a first conductivity to a second conductivity, that increases at a second rate from the second conductivity to a third conductivity, and that increases at a third rate from the third conductivity to a fourth conductivity; and
(c) eluting the antibody from the ion exchange material, after the washing step (b).