Method for protein purification
AssigneeUcb Biopharma Srl
InventorsSAM PHILIP HEYWOOD, GAVIN BARRY WILD
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Abstract
The present invention provides a method for recovering a human VH3 domain-containing antibody in monomeric form. In particular the present invention provides a new method that allows recovery of monomeric human VH3 domain-containing antibodies from a mixture containing monomeric and multimeric forms of the antibody.
Claims10
A method for obtaining a human VH3 domain-containing antibody in monomeric form comprising:
- a) applying a mixture comprising a human VH3 domain-containing antibody in monomeric and multimeric form to a protein A chromatography material wherein said protein A comprises domain D and/or E, under conditions that allow binding of said antibody to protein A, and
- b) recovering the human VH3 domain-containing antibody in monomeric form,
- wherein the human VH3 domain-containing antibody does not contain an Fc region and said VH3 domain-containing antibody comprises SEQ ID NOs:
- 11 and 12 or SEQ ID NOs:
- 29 and 31.
A method for manufacturing a human VH3 domain-containing antibody comprising:
- a) expressing the antibody in a host cell,
- b) recovering a mixture containing the antibody, host cells and other contaminants,
- c) purifying the antibody using at least a protein A chromatography step wherein said protein A comprises domain D and/or E, and
- d) recovering the human VH3 containing antibody,
- wherein the human VH3 domain containing antibody does not contain an Fc region and said VH3 domain-containing antibody comprises SEQ ID NOs:
- 11 and 12 or SEQ ID NOs:
- 29 and 31.
A method of separating a human VH3 domain-containing antibody in monomeric form from the antibody in multimeric form comprising:
- a) applying a mixture comprising a human VH3 domain-containing antibody in monomeric and multimeric form to a protein A chromatography material wherein said protein A comprises domain D and/or E,
- b) allowing binding of said antibody to protein A,
- c) applying an elution buffer that selectively disrupts binding of the antibody in monomeric form,
- d) recovering the resulting eluate, and optionally
- e) applying a second elution buffer that disrupts binding of the antibody in multimeric form and recovering this second eluate,
- wherein the human VH3 domain-containing antibody does not contain an Fc region and said VH3 domain-containing antibody comprises SEQ ID NOs:
- 11 and 12 or SEQ ID NOs:
- 29 and 31.
A method of separating a human VH3 domain-containing antibody in monomeric form from the antibody in multimeric form comprising:
- a) applying a mixture comprising a human VH3 domain-containing antibody in monomeric and multimeric form to a protein A chromatography material wherein said protein A comprises domain D and/or E,
- b) allowing binding of the antibody in multimeric form,
- c) recovering the antibody in monomeric form in the flow-through, and optionally
- d) applying an elution buffer that selectively disrupts binding of the antibody in multimeric form, and
- e) recovering the eluate resulting from d);
- wherein the human VH3 domain-containing antibody does not contain an Fc region and said VH3 domain-containing antibody comprises SEQ ID NOs:
- 11 and 12 or SEQ ID NOs:
- 29 and 31.
Dependencies are inferred from the claim text.