Drug – bio-affecting and body treating compositions – Immunoglobulin – antiserum – antibody – or antibody fragment,... – Monoclonal antibody or fragment thereof
Patent
1997-01-23
2000-11-28
Kunz, Gary L.
Drug, bio-affecting and body treating compositions
Immunoglobulin, antiserum, antibody, or antibody fragment,...
Monoclonal antibody or fragment thereof
435325, 435334, 5303881, A61K 39395, C12N 512, C12P 2108, C07K 1628
Patent
active
061531901
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention relates generally to the generation of monoclonal antibodies by use of a selected antigen: and more particularly to the generation of monoclonal antibodies which are receptor agonists.
BACKGROUND OF THE INVENTION
A vast majority of receptors of the single transmembrane class respond to ligand binding by some form of aggregation. This aggregation can be between identical receptor subunits (as in homodimerization, homotrimerization, etc.) or between different receptor subunits (as is heterodimerization, heterotrimerization, etc). This aggregation appears to be part of the signal for the target cell to respond biologically, in that mutants of the ligand which are unable to interact with the second subunit are still able to bind, but no longer cause dimerization and biological activation of the receptor [P. R. Young, Curr. Opin. Biotech., 3:408-421 (1992)].
For example, there is evidence in the literature that suggests dimerization of the erythropoietin receptor (EpoR) upon ligand binding [S. S. Watowich et al., Molec. Cell Biol., 14:3535-3549 (1992) and S. S. Watowich et al., Proc. Natl. Acad. Sci., USA, 89:2140-2144 (1992)]. Reports about IL-6 have indicated that its second subunit gp130 may dimerize upon IL-6 binding [M. Murkami et al., Science, 260:1808-1810 (1993)]. for some receptors in which homodimerization is induced by ligand binding, monoclonal antibodies (mAbs) were discovered which had agonist properties. These include mAbs to EGF, TNF and growth hormone receptors [A. B. Schrieber et al., J. Biol. Chem., 258:846-853 (1983); L. H. K. Defize et al., EMBO J., 5:1187-1192 (1986); H. Engelmann et al., J. Biol. Chem., 265:14497-14505 (1990); and G. Fuh et al., Science, 256:1677-1680 (1992)]. In each case, these mAbs, by virtue of their two antigen recognition sites, were able to bring together two receptors and thus activate them. Fab fragments made from these mAbs were inactive. In some cases, the apparent affinity of the antibody for receptor was comparable to that of the ligand, e.g., growth hormone [Fuh et al., cited above].
It has also been discovered that antibodies to IL-3 receptor have agonist properties [Suguwara et al., J. Immunol., 140:526-530 (1988)]. Previous literature has described the production of anti-erythropoietin receptor antibodies [A. D'Andrea et al., Blood, 82:46-52 (1993); A. D'Andrea et al., Blood, 84:1982-1991 (1994) and M-G Yet et al., Blood, 82: 1713-1719 (1993). See also, PCT Application WO96/03438 published Feb. 8, 1996. While the Yet et al., reference suggests the occurrence of possible EPO-like activity in one mAb, the mAb is not characterized. Neither Yet et al., nor the other literature provides any reproducible manner of generating agonist mAbs.
There remains a need in the art for the development of additional mAbs which have an affinity for receptors comparable to that of the ligand, and which can act as agonists of the receptor.
SUMMARY OF THE INVENTION
In one aspect, the present invention provides a method for reliably generating an antibody which is an agonist of a receptor. This method employs as the immunizing antigen, a recombinant immunogen which consists of a first extracellular domain of a receptor molecule spaced apart from a second extracellular domain from that receptor by a bridging moiety. The bridging moiety places the first domain and the second domain into a functional proximity which mimics the functional domain orientation and proximity of the naturally occurring multimeric receptor. The bridging moiety can be an amino acid spacer peptide, an organic molecule, an Fe portion of a human immunoglobulin or an amphipathic helix, for example.
In another aspect, the invention includes antibodies produced by the above-described method. The antibodies so generated are characterized by the ability to bind to the naturally occurring receptor and by such binding initiate the biological activity of the receptor. The antibodies of the invention may be chimeric antibodies, humanized antibodies, monoclonal antibo
REFERENCES:
Callan et al., PNAS USA vol. 90, pp. 10454-10458, 1993.
J. Immunol., vol. 142, No. 3, Feb. 1, 1989, Schreurs et al., A monoclonal antibody with IL-3 like acitivity blocks IL-3 binding and stimulates tyrosine phosphorylation pp. 819-825.
Blood, vol. 82, No. 6, Sep. 15, 1993 Yet et al. The extracytoplasmic domain of the erythropoietin receptor forms a monomeric complex with erythropoietin pp.1713-1719.
Science, vol. 256, Jun. 19, 1992 Fuh et al. Rational design of potent antagonist to the human growth hormone receptor pp.1677-1680.
Proc. Natl. Acad. Sci USA vol. 89 Mar. 1992 Watowich et al. Homodimerization and constitutive activation of the erythropoientin receptor pp.2140-2144.
Blood, vol. 82, No. 1, Jul. 1, 1993 D'Andrea et al. Anti-erthropoientin receptor antibodies inhibit erythropoietin binding and neutralize bioactivity pp. 46-52.
J. Immunol., vol. 140, No. 2, Jan., 1992, Sugawara et al., monoclonal antibodies with interleukin-3 like acitivity derived from a MRL/lpr mouse pp. 526-530.
Erickson-Miller Connie L.
Young Peter Ronald
Baumeister Kirk
Gucker Stephen
King William T.
Kunz Gary L.
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