Mammalian cytokines; related reagents and methods
View Patent ↗Purified genes encoding cytokine from a mammal, reagents related thereto including purified proteins, specific antibodies, and nucleic acids encoding this molecule are provided. Methods of using said reagents and diagnostic kits are also provided.
1. A method for preparing an antibody or a fragment thereof that comprises the antigen binding site of the antibody, wherein said antibody binds to:
(i) a mammalian IL-B30/p40 complex comprising an IL-B30 subunit (SEQ ID NO:2 or 4) and a p40 subunit (SEQ ID NO:6 or 8), or
(ii) a fusion protein comprising the IL-B30 subunit (SEQ ID NO:2 or 4) and the p40 subunit (SEQ ID NO:6 or 8);
said method consists of:
(a) raising an antibody to said IL-B30/p40 complex or a polypeptide that comprises at least 11 contiguous amino acids from said p40 subunit and at least 11 contiguous amino acids from the IL-B30 subunit,
(b) confirming that said antibody binds to the IL-B30/p40 complex or fusion protein; and
(c) optionally preparing said fragment from said antibody.
2. The method of claim 1 , wherein said antibody or fragment thereof is a humanized antibody, monoclonal antibody, single chain antibody, Fv fragment, Fab fragment, Fab′ fragment, or F(ab′)2 fragment.
3. The method of claim 1 , wherein said antibody is a neutralizing antibody.
4. The method of claim 1 , wherein the fusion protein comprises, from the N-terminus to the C-terminus, mature IL-12 p40, a linker, and mature IL-B30.
5. A method of selecting an antibody or fragment thereof that specifically binds to:
(i) a mammalian IL-B30/p40 complex comprising an IL-B30 subunit (SEQ ID NO:2 or 4) and a p40 subunit (SEQ ID NO:6 or 8), or
(ii) a fusion protein comprising the IL-B30 subunit (SEQ ID NO:2 or 4) and the p40 subunit (SEQ ID NO:6 or 8);
wherein the antibody or fragment thereof does not bind the IL-B30 or p40 subunit alone, said method comprising
(a) providing candidate antibodies or fragments thereof that bind to the IL-B30/p40 complex or fusion protein; and
(b) detecting if the candidate antibodies or fragments thereof bind the IL-B30 or p40 subunit, and selecting the antibody or fragment thereof that does not bind the IL-B30 or p40 subunit.
6. The method of claim 5 wherein said antibody or fragment thereof is selected by immunoselection or immunodepletion.
7. The method of claim 5 , wherein said antibody or fragment thereof is a humanized antibody, monoclonal antibody, single chain antibody, Fv fragment, Fab fragment, Fab′ fragment, or F(ab′)2 fragment.
8. The method of claim 5 , wherein said antibody is a neutralizing antibody.
9. The method of claim 5 , wherein the fusion protein comprises, from the N-terminus to the C-terminus, mature IL-12 p40, a linker, and mature IL-B30.
10. The method of claim 5 , wherein said antibody or fragment thereof is detectably labeled.