Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1985-11-18
1989-06-20
Warden, Robert J.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 18, 435 21, 435 26, 435 28, 436500, 436507, 436512, 436518, 436536, 436548, 436800, 436804, 436805, 436808, 935110, 530387, 530388, 530808, G01N 3353
Patent
active
048408956
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to a class of monoclonal antibodies, which may be free or present in a complex, to the manufacture of such antibodies and to their use in reactions which employ an antigen-antibody reaction.
Reactions between an antigen and its antibody have found many applications in biotechnology and especially in diagnostic tests, for example medical diagnostic tests, gene probes and the like. Monoclonal antibodies (see for example G. Galfre and C. Milstein, Methods in Enzymology, 73, 3-57, 1981) have been found to be of particular use in biotechnology and especially in diagnostic tests.
A monoclonal antibody was described by D. A. Nemazee and V. J. Sato (see Proc. Natl. Acad. Sci. USA, V2, 79, pp 3828-3832, 1982) which was an antibody against a complex of two other antibodies as a result of exposing a new epitope in the Fc region of one of the original antibodies. Since the original antibodies were both macromolecules conformational changes in the Fc region was not unexpected. Similar changes would not be expected to occur if a small molecule had been employed. Similarly since small molecules cannot have simultaneous multiple epitopes and cannot thus themselves bind more than one antibody, (for example see R. J. Thompson and A. P. Jackson, TIBS, 9, pp1-3, 1984) the effect described by Nemazee and Sato would not be expected if a complex employing a small molecule was used. Contrary to this expectation, it has now been found that, possibly because of a different mechanism, a new class of monoclonal antibody can be made which are surprisingly useful in biotechnology, for example in diagnostic tests.
Accordingly the present invention provided a monoclonal antibody characterised in that said monoclonal antibody is a secondary monoclonal antibody against a complex of a small molecule and a binding protein against said small molecule which secondary antibody is not an antibody against the small molecule or against its binding protein.
When used herein the term "small molecule" means a molecule of molecular weight less than 5000. Such small molecules most aptly have a molecular weight of less than 2000 and preferably have a molecular weight of less than 1200.
Proteins used in this invention may be antibodies or other proteinaceous materials such as enzymes, binding proteins such as steroid binding proteins or vitamin binding proteins. Preferably the protein used in this invention is a monoclonal antibody in which case the small molecule may be termed herein a small antigen.
The skilled art worker will appreciate that low molecular weight materials such as the small molecules used in this invention are normally non-immunogenic but that antibodies thereto can be obtained by immunising an animal with a conjugate of the non-immunogenic molecule (or a very close analogue) and an immunogenic material such as bovine serum albumin or an equivalent agent. The desired antibody may then be obtained by methods known per se.
Accordingly, in a preferred form, the present invention provides a monoclonal antibody characterised in that said monoclonal antibody is a secondary monoclonal antibody against a complex of a small antigen and a monoclonal antibody against the small antigen which secondary monoclonal antibody is not an antibody against the small antigen or against its monoclonal antibody.
When used hereinafter the term "secondary monoclonal antibody" will mean an antibody provided by this invention as defined above. When used hereinafter the term "primary monoclonal antibody" will mean the monoclonal antibody against the antigen (that is a conventional monoclonal antibody).
The monoclonal antibody may be the complete immunoglobulin or fragment thereof having the described binding activity. As will become apparent hereinafter, certain embodiments of this invention will benefit more from the use of the complete immunoglobulin than from a fragment thereof whereas other embodiments will benefit more from the use of fragments of the complete immunoglobulin such as the Fab and F(ab').sub.2 fragments. This reflec
REFERENCES:
patent: 4510239 (1985-04-01), Miller et al.
patent: 4544640 (1985-10-01), Soma et al.
patent: 4606855 (1986-08-01), Deutsch et al.
patent: 4670383 (1987-06-01), Baier et al.
D. A. Nemazee et al., Proc. Natl. Acad. Sci. USA, 79, pp. 3828-3832, Jun., 1982.
Science, vol. 218, Oct. 29, 1982, p. 474.
Benson Robert
Cambridge Patent Development Limited
Warden Robert J.
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