Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1993-05-10
1998-12-15
Elliott, George G.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
4351723, 4353201, G01N 3353, C12N 1564, C12N 1570
Patent
active
058495002
DESCRIPTION:
BRIEF SUMMARY
This application is a 371 of PCT/EP92/01524, filed Jul. 6, 1992.
The present invention concerns phagemids for the selection of specific antibodies from large recombinant libraries, the production of these phagemids and their use to select specific antibodies from large recombinant libraries using small amounts of antigen.
Plasmid and phage antibody libraries have been established in E. coli from PCR amplified immunoglobulin families following immunization. Recombinant antibodies to immunogens were selected by an ELISA assay of the bacterial supernatant from isolated bacterial colonies (Ward, E. S., Gussow, D., Griffiths, A. D., Jones, P. T. & Winter, G.: Binding activities of a repertoire of single immunoglobulin variable domains secreted from Escherichia coli. Nature 341 (1989) 544-546) or by screening nitrocellulose plaque lift-offs of bacterial colonies for reactivity to the radioactively labeled immunogen (Huse, W. D., Sastry, L., Iverson, S. A., Kang, A. S., Alting-Mees, M., Burton, D. R., Benkovic, S. J. and Lerner, R. A.: Generation of a large combinatorial library of the immunoglobin repertoire in phage lambda. Science 246 (1989) 1275-1281). However, for the selection of specific antibodies from randomly combined light and heavy chain libraries of non-immunized animals that do not contain a preponderance of antibodies to a particular antigen, a procedure is required for screening millions of antibody producing bacteria.
A possible way to screen a broad range of antibodies is to attach recombinant antibodies to the surface of bacteria or bacteriophages so that they can then be rapidly selected by antigens bound to a solid phase. Given the difficulties of targeting proteins to the cell surface of bacteria, an attractive candidate in view of its small size and relatively simple genetic make-up is the M13 family of filamentous bacteriophages (for reviews see Webster, R. E. and Lopez, J. in "Virus Structure and Assembly" ed. S. Casjens, publ. Jones and Bartlett Inc., Boston/Portala Valley, USA, 1985; Day, L. A., Marzec, C. J., Reisberg, S. A. and Casadevall, A.: DNA packaging in filamentous bacteriophages. Ann. Rev. Biophys. Biophys. Chem. 17 (1988) 509-539).
The product of gene III (pIII) is a relatively flexible and accessible molecule composed of two functional domains; an amino-terminal domain that binds to the F pilus of male bacteria during infection and a carboxy-terminal domain buried within the virion that is important for morphogenesis. Peptides can be inserted between the two domains of pIII (Smith, G. P.: Filamentous fusion phage: novel expression vectors that display cloned antigens on the virion surface. Science 228 (1985) 1315-1317) or near the N-terminus (Parmley, S. F. and Smith, G. P.: Antibody-selectable filamentous fd phage vectors: affinity purification of target genes. Gene, 73 (1988) 305-318) without destroying its functions in morphogenesis and infection. After much pioneering work on the use of pIII in fd phages for carrying foreign peptides, Parmely and Smith (1988, a.a.O.) showed that peptide epitopes inserted at the aminoterminal end could bind phages to immobilized antibodies. As a consequence of this work it has been possible to generate peptide libraries that can be screened for binding to ligands and antibodies (Scott, J. K. and Smith, G. P.: Searching for peptide ligands with an epitope library. Science 249 (1990) 386-390; Devlin, J. J., Panganiban, L. C. and Devlin, P. E.: Random peptide libraries: A source of specific protein binding molecules. Science 249 (1990) 404-406; Cwirla, S. E., Peters. E. A., Barrett, R. W. and Dower, W. J. Peptides on phage, a vast library of peptides for identifying ligands.: Proc. Natl. Acad. Sci. USA, 87 (1990) 6378-6382).
McCafferty, J., Griffiths, A. D., Winter, G. and Chiswell, D. J.: Phage antibodies: filamentous phage displaying antibody variable domains. Nature, 348 (1990) 552-554 reported the assembly of an antibody-pill fusion protein into an fd phage with a Tet.sup.R gene after inserting antibody DNA into the 5' end of gene III. T
REFERENCES:
Muller, H., Ph.D. thesis, Univ. of Heidelberg, cover sheets and pp. 56-58 989).
McCafferty et al, Nature 348, pp. 552-554 (1990).
Ward et al, Nature 341, pp. 544-546 (1989).
Cwirla et al, Proc. Natl. Acad. Sci. USA 87, pp. 6378-6382 (1990).
Scott et al, Science 249, pp. 386-390 (1990).
Breitling et al, Gene 104, pp. 147-153 (1991).
Huse et al, Science 246, pp. 1275-1281 (1989).
Day et al, Ann. Rev. Biophys. Biophys. Chem. 17, pp. 509-539 (1988).
Smith et al, Science 228, pp. 1315-1317 (1985).
Parmley et al, Gene 73, pp. 305-318 (1988).
Devlin et al, Science 249, pp. 404-406 (1990).
Amit et al, Science 233, pp. 747-753 (1986).
Verhoeyen et al, Science 239, pp. 1534-1536 (1988).
Breitling et al, J. Mol. Biol. 189, pp. 367-370 (1986).
Bass et al, Proteins: Structure, Function and Genetics, vol. 8, No. 4 1990, Wiley Press, NY, US; pp. 309-314.
Bujard et al, Methods Enzymol. 155, pp. 416-433 (1987).
Lanzer et al. Proc. Natl. Acad. Sci. USA 85, pp. 8973-8977 (1988).
Goldsmith et al, Biochemistry 16, pp. 2686-2694 (1977).
Boeke et al, Proc. Natl. Acad. Sci. USA 79, pp. 5200-5204 (1982).
Crissman et al, Virology 132, pp. 445-455 (1984).
Davis et al, J. Mol. Biol. 181, pp. 111-121 (1985).
Pluckthun et al, J. Biol. Chem. 262, 3951-3957 (1987).
Laemmli et al, Nature 227, pp. 680-685 (1970).
Towbin et al, Proc. Natl. Acad. Sci. USA 76, pp. 4350-4354 (1979).
Webster et al., Structure and Assembly of the Class I Filamentous Bacteriophage, Duke University Medical Center pp. 236-267 (1985).
Sambrook et al, "Molecular Cloning", 1989, pp. 4.17-4.19 and 17.29-17.30.
Braunagel Michael
Breitling Frank
Dubel Stefan
Klewinghaus Iris
Little Melvyn
Deutsches Krebsforschungszentrum Stiftung des Offentlichen Recht
Elliott George G.
McKelvey Terry A.
LandOfFree
Phagemid for antibody screening does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Phagemid for antibody screening, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Phagemid for antibody screening will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1456547