Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1986-02-07
1989-05-09
Warden, Robert J.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 21, 435810, 436506, 436507, 436513, 436518, 436547, 436805, 436808, 530387, G01N 3353
Patent
active
048289850
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to a class of polyclonal antibodies, which may be free or present in a complex, to the preparation of such antibodies and to their use in reactions which employ antibody-antigen interactions.
Reactions between an antigen and its antibody have found many applications in biotechnology and especially in diagnostic tests, for example medical (including veterinary) diagnostic tests, gene probes, and the like. Recently the art has moved forward rapidly due in large part to the introduction of monoclonal antibodies which allow great specificity and accuracy (for a discussion of monoclonal antibodies see for example G. Galfre and C. Milstein, Methods in Enzymology, 73, 3-57, 1981).
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, pp 1-3, 1984) the effect described by Nemazee and Sato would not be expected if a complex employing a small molecule was used.
I have now discovered that, if the normal tendency to use monoclonals is reversed, certain new polyclonal antibodies may be produced by a simple process and be used in reactions which employ antibody-antigen interactions. I have found that if certain polyclonal antibodies are made free of certain other antibodies, then they are of use in, inter alia, diagnostic tests and are particularly advantageous in such tests if they are labelled.
Accordingly the present invention provides a polyclonal antibody characterised in that said polyclonal antibody is a secondary polyclonal antibody against a complex of a non-immunogenic molecule and a binding protein against said non-immunogenic molecule which secondary antibody is free from antibodies againt the non-immunogenic molecule and against its binding protein.
The non-immunogenic molecule is normally a small molecule, that is a molecule of molecular weight less than 5000. Such small molecules more aptly have a molecular weight of less than 2000 and favourably have a molecular weight of less than 1200.
The skilled art worker will appreciate that low molecular weight materials 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) with an immunogenic material such as bovine serum albumin or an equivalent agent. The desired antibody may then be obtained by methods known per se.
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 an antibody in which case the small molecule may be termed herein a small antigen.
Thus in a favoured aspect the present invention provides a secondary polyclonal antibody against a complex of a non-immunogenic molecule and a primary antibody against said non-immunogenic molecule said secondary polyclonal antibody being free from antibodies against the non-immunogenic molecule and its primary antibody.
Herein the term "primary antibody" refers to an antibody which will bind the non-immunogenic molecule per se. This primary antibody may be a polyclonal antibody or a monoclonal antibody. It is desirable to employ a polyclonal antibody because they are more readily available and surprisingly it has been found to be effective when producing the secondary antibody of this invention. It is also sometimes desirable to employ a monoclonal primary antibody since, although more di
REFERENCES:
patent: 4132767 (1979-01-01), Tohmatsu et al.
patent: 4544640 (1985-10-01), Soma et al.
patent: 4670383 (1987-06-01), Baier
International Search Report, Nemazee, D. et al: "Enchancing Antibody: A Novel Component of the Immune Response", pp. 3828-3822.
Chemical Abstract, vol. 92, No. 9, 3 Mar. 1980, p. 483, Abstract 74158d.
Chemical Abstract, vol. 83, No. 1, 7 Jul. 1975, p. 600, Abstract 69335.
Chemical Abstract, vol. 95, No. 1, 6 Jul. 1981, p. 487, Abstract 49285.
Benson Robert
Cambridge Patent Developments Limited
Warden Robert J.
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