Labelling and selection of molecules

• US 6,342,588 B1
• Filed: 08/16/1999
• Issued: 01/29/2002
• Est. Priority Date: 07/08/1996
• Status: Expired due to Term

• Current Assignee: Cambridge Silicon Radio Limited (Qualcomm, Inc.)
• Original Assignee: Cambridge Silicon Radio Limited (Qualcomm, Inc.)
• Inventors: Johnson, Kevin Stuart, McCafferty, John Gerald, Osbourn, Jane Katharine, Derbyshire, Elaine Joy, Vaughan, Tristan John
• Primary Examiner(s): BRUMBACK, BRENDA G
• Application Number:
  US09/375,314
• Time in Patent Office: 897 Days
• Field of Search: 530/412, 530/413, 435/5, 435/6, 435/7.1, 435/7.2, 435/7.9
• US Class Current: 530/413
• 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: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 1/36 by a combination of two or more processes of different types
• CPC Class Codes: C07K 14/523 Beta-chemokines, e.g. RANTES, I-309/TCA-3, MIP-1alpha, MIP-1beta/ACT-2/LD78/SCIF, MCP-1/MCAF, MCP-2, MCP-3, LDCF-1, LDCF-2
• CPC Class Codes: C07K 14/70542 CD106
• CPC Class Codes: C07K 14/70596 Molecules with a "CD"-designation not provided for elsewhere
• CPC Class Codes: C07K 14/82 Translation products from oncogenes
• 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 16/065 Purification, fragmentation
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 16/2854 against selectins, e.g. CD62
• CPC Class Codes: C07K 16/2866 against receptors for cytokines, lymphokines, interferons
• CPC Class Codes: C07K 16/2896 against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C07K 16/3007 Carcino-embryonic Antigens
• CPC Class Codes: C07K 16/32 against translation products of oncogenes
• CPC Class Codes: C07K 16/4266 against anti-tumor receptor Ig
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C12N 9/16 acting on ester bonds (3.1)
• CPC Class Codes: G01N 33/532 Production of labelled immunochemicals
• CPC Class Codes: G01N 33/535 : with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates

Abstract

A method of labelling molecules which includes providing in a common medium a label molecule, a marker ligand able to bind a member of a specific binding pair, such as an antigen, a sbp member, an enzyme able to catalyse binding of the label molecule to other molecules, the enzyme being associated with the marker ligand; causing or allowing binding of the marker ligand to the sbp member; and causing or allowing binding of the label molecule to other molecules in the vicinity of the marker ligand bound to the sbp member. The marker ligand may be an antibody or any specific binding molecule, such as a chemokine or cytokine. A complementary member of the specific binding pair may be included, e.g. an antibody, or a diverse population of such sbp members, e.g. antibodies, may be included within which those which bind the counterpart sbp member, e.g. antigen, may be labelled and subsequently isolated for manipulation and/or use. Suitable labels include biotin-tyramine with signal transfer being catalysed by hydrogen peroxidase. Cells, virus particles and other moieties may be labelled, for identification or obtention of proteins which interact or are in close proximity with a particular sbp member, or of cells of interest, or for enhancement of labelling, e.g. for cell sorting.

Claims

1. A method of obtaining a peptide or polypeptide, the method comprising

providing in a common medium:

label molecules;

a marker ligand able to bind a member of a specific binding pair (sbp), which specific binding pair has a first sbp member and a second sbp member, and wherein said marker ligand is able to bind said second sbp member;

said second sbp member;

an enzyme able to catalyze binding of said label molecules to proteins, peptides or ligands, said enzyme being conjugated to said marker ligand;

causing or allowing binding of said marker ligand to said second sbp member; and

causing or allowing binding of said label molecules to proteins, peptides or ligands in the vicinity of said marker ligand bound to said second sbp member thereby providing labeled proteins, peptides or ligands;

wherein said proteins, peptides or ligands to which said label molecules bind comprise a sbp member (“

first sbp member”

) which binds said second sbp member or comprise a sbp member (“

further sbp member”

) which binds a protein, peptide or ligand in the vicinity of said second sbp member, and said first sbp member or said further sbp member is displayed on the surface of a virus particle;

the method further comprising;

isolating and/or purifying said particle from the medium following binding of said first sbp member to said second sbp member or binding of said further sbp member to a protein, peptide or ligand in the vicinity of said second sbp member, and labeling of said particle and/or said displayed first or further sbp member;

obtaining nucleic acid from an isolated and/or purified particle; and

producing by expression from nucleic acid with the sequence of said nucleic acid obtained from said particle or a sequence modified by the addition, insertion, deletion and/or substitution of one or more nucleotides an encoded peptide or polypeptide.

2. A method according to claim 1 wherein said particle is isolated and/or purified by means of a member of a specific binding pair able to bind said labeled particle and/or displayed first or further sbp member.

3. A method according to claim 1 wherein the peptide or polypeptide produced by expression from said nucleic acid is formulated into a composition including at least one additional component.

4. A method according to claim 3 wherein the composition includes a pharmaceutically acceptable vehicle or carrier.