Altered polypeptides with increased half-life

• US 5,869,046 A
• Filed: 04/14/1995
• Issued: 02/09/1999
• Est. Priority Date: 04/14/1995
• Status: Expired due to Term

• Current Assignee: Genentech, Inc. (Roche Holding AG)
• Original Assignee: Genentech, Inc. (Roche Holding AG)
• Inventors: Presta, Leonard G., Snedecor, Bradley R.
• Primary Examiner(s): Eisenschenk, Frank C.
• Assistant Examiner(s): RABIN, EVELYN MARIE
• Application Number:
  US08/422,092
• Time in Patent Office: 1,397 Days
• Field of Search: 435/69.7, 435/172.1, 530/387.3, 530/388.15, 530/391.1, 424/133.1, 424/153.1, 935/15
• US Class Current: 424/133.1
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/2845 against integrin beta2-subunit-containing molecules, e.g. CD11, CD18
• CPC Class Codes: C07K 2319/00 Fusion polypeptide

Abstract

Polypeptides that are cleared from the kidney and do not contain in their original form a Fc region of an IgG are altered so as to comprise a salvage receptor binding epitope of an Fc region of an IgG and thereby have increased circulatory half-life.

Claims

1. A method for preparing a variant polypeptide of interest, wherein the polypeptide of interest is a Fab or a (Fab'"'"')₂ which contains an Ig domain or Ig-like domain comprising at least one of a CH1 or C_L, region, that is cleared from the kidney and does not contain a Fc region of an IgG, comprising:

altering said polypeptide of interest within the CH1 or C_L, region to contain a salvage receptor binding epitope taken from two loops of a CH2 domain of an Fc region of an IgG, and wherein said altering results in an increased in vivo half-life.

2. The method of claim 1 wherein the altering step is conducted by site-directed, cassette, or PCR mutagenesis.

3. The method of claim 1 wherein the polypeptide of interest is a Fab, or (Fab'"'"')₂, of an anti-LFA-1 antibody.

4. The method of claim 3 wherein the polypeptide of interest is an anti-CD18 Fab or anti-CD18 (Fab'"'"')₂.

5. The method of claim 4 wherein the polypeptide of interest is human or humanized.

6. The method of claim 1 wherein the epitope is fused to the polypeptide of interest.

7. A method for preparing a variant polypeptide of interest, wherein the polypeptide of interest is an anti-CD18 Fab or anti-CD18 (Fab'"'"')₂ which contains an Ig domain or Ig-like domain comprising at least one of a CH1 or C_L, region, that is cleared from the kidney and does not contain a Fc region of an IgG, comprising:

altering said polypeptide of interest within the CH1 or C_L region to contain a salvage receptor binding epitope taken from two loops of a CH2 domain of an Fc region of an IgG, and wherein said altering results in an increased in vivo half-life.