Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Blood proteins or globulins – e.g. – proteoglycans – platelet...
Patent
1993-01-08
1999-10-19
Eisenschenk, Frank C.
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
Blood proteins or globulins, e.g., proteoglycans, platelet...
530867, 435 91, 435 691, 435 698, 4353201, 4352351, 536 231, 536 2353, 536 234, C07K 1900, C07H 2104, C12N 1513
Patent
active
059691085
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to methods for producing members of specific binding pairs. The present invention also relates to the biological binding molecules produced by these methods.
Owing to their high specificity for a given antigen, the advent of monoclonal antibodies (Kohler, G. and Milstein C; 1975 Nature 256: 495) represented a significant technical break-through with important consequences both scientifically and commercially.
Monoclonal antibodies are traditionally made by establishing an immortal mammalian cell line which is derived from a single immunoglobulin producing cell secreting one form of a biologically functional antibody molecule with a particular specificity. Because the antibody-secreting mammalian cell line is immortal, the characteristics of the antibody are reproducible from batch to batch. The key properties of monoclonal antibodies are their specificity for a particular antigen and the reproducibility with which they can be manufactured.
Structurally, the simplest antibody (IgG) comprises four polypeptide chains, two heavy (H) chains and two light (L) chains inter-connected by disulphide bonds (see FIG. 1). The light chains exist in two distinct forms called kappa (K) and lambda (.lambda.). Each chain has a constant region (C) and a variable region (V). Each chain is organized into a series of domains. The light chains have two domains, corresponding to the C region and the other to the V region. The heavy chains have four domains, one corresponding to the V region and three domains (1,2 and 3) in the C region. The antibody has two arms (each arm being a Fab region), each of which has a VL and a VH region associated with each other. It is this pair of V regions (VL and VH) that differ, from one antibody to another (owing to amino acid sequence variations), and which together are responsible for recognising the antigen and providing an antigen binding site (ABS). In even more detail, each V region is made up from three complementarity determining regions (CDR) separated by four framework regions (FR). The CDR's are the most variable part of the variable regions, and they perform the critical antigen binding function. The CDR regions are derived from many potential germ line sequences via a complex process involving recombination, mutation and selection.
It has been shown that the function of binding antigens can be performed by fragments of a whole antibody. Example binding fragments are (i) the Fab fragment consisting of the VL, VH, CL and CH1 domains; (ii) the Fd fragment consisting of the VH and CHI domains; (iii) the Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (iv) the dAb fragment (Ward, E. S. et al., Nature 341, 544-546 (1989) which consists of a VH domain; (v) isolated CDR regions; and (vi) F(ab').sub.2 fragments, a bivalent fragment comprising two Fab fragments linked by a disulphide bridge at the hinge region.
Although the two domains of the Fv fragment are coded for by separate genes, it has proved possible to make a synthetic linker that enables them to be made as a single protein chain (known as single chain Fv (scFv); Bird, R. E. et al., Science 242, 423-426 (1988) Huston, J. S. et al., Proc. Natl. Acad. Sci., USA 85, 5879-5883 (1988)) by recombinant methods. These scFv fragments were assembled from genes from monoclonals that had been previously isolated. In this application, the applicants describe a process to assemble scFv fragments from VH and VL domains that are not part of an antibody that has been previously isolated.
Whilst monoclonal antibodies, their fragments and derivatives have been enormously advantageous, there are nevertheless a number of limitations associated with them.
Firstly, the therapeutic applications of monoclonal antibodies produced by human immortal cell lines holds great promise for the treatment of a wide range of diseases (Clinical Applications of Monoclonal Antibodies. Edited by E. S. Lennox.
British Medical Bulletin 1984. Publishers Churchill Livingstone). Unfortunately, immortal antibody-produc
REFERENCES:
patent: 4593002 (1986-06-01), Dulbecco
patent: 4816397 (1989-03-01), Boss et al.
patent: 4946778 (1990-08-01), Ladner et al.
patent: 5091513 (1992-02-01), Huston et al.
patent: 5223409 (1993-06-01), Ladner et al.
patent: 5403484 (1995-04-01), Ladner et al.
patent: 5427908 (1995-06-01), Dower et al.
Bass et al., Proteins, Structure, Function and Genetics 8:309-314 (1990).
de la Cruz et al., J. Biol. Chem. 263(9):4318-4322 (1988).
Huse et al., Science 246:1275-1281 (1989).
Kang et al., Proc. Natl. Acad. Sci. USA 88:4363-4366 (1991).
McCafferty et al., Nature 348:552-554 (1990).
Milstein, F.R.S., Proc. R. Soc. London B 239:1-16 (1990).
Parmley and Smith, Gene 73:305-318 (1988).
Short et al., Nucleic Acids Research 16(15):7583-7600 (1988).
Smith, Science 228:1315-1317 (1985).
Tsunetsugu-Yokota et al., Gene 99:261-265 (1991).
Winter and Milstein, Nature 349:293-299 (1991).
Kabat Sequences of Proteins of Immunological Interest 4th ed. (1987), p. 181.
Sambrook et al. Molecular Cloning: A Laboratory Manual 4.13, 4.14, 4.17-4.19, 4.46 and A.12 (1989).
Bonnert Timothy Peter
Chiswell David John
Clackson Timothy Piers
Griffiths Andrew David
Holliger Kaspar Philipp
Cambridge Antibody Technology Limited
Eisenschenk Frank C.
Medical Research Council
LandOfFree
Methods for producing members of specific binding pairs does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Methods for producing members of specific binding pairs, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Methods for producing members of specific binding pairs will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2056908