Expression vectors encoding bispecific fusion proteins and metho

Details Expression vectors encoding bispecific fusion proteins and metho Expression vectors encoding bispecific fusion proteins and metho

1995-10-05

2000-10-17

Guzo, David

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

4353201, 435326, 435328, 5303873, 5303879, C12P 2102, C12N 1585, C07K 1628

Patent

active

061329924

ABSTRACT:
The present invention provides an expression vector encoding monospecific or bispecific fusion protein. In one embodiment the expression vector encodes a monospecific fusion protein, which vector comprises a recombinant monospecific single chain cassette comprising a DNA sequence encoding a first binding domain capable of binding a cell surface antigen. In another embodiment the expression vector encodes a bispecific fusion protein, which vector comprises a recombinant bispecific single chain cassette comprising a DNA sequence encoding a first binding domain capable of binding a cell surface antigen and a DNA sequence encoding a second binding domain capable of binding a cell surface antigen, each domain capable of binding a different antigen. The present invention also provides a method for producing a biologically active monospecific or bispecific fusion protein in a mammalian cell. This method comprises: (a) transfecting the mammalian cell with the recombinant expression vector of the invention; (b) culturing the mammalian cell so transfected in step (a); and (c) recovering the biologically active bispecific fusion protein so as produced by the cultured mammalian cell.

REFERENCES:
patent: 5637481 (1997-06-01), Ledbetter et al.
Adams, GP et al., "Highly specific in vivo tumor targeting by monovalent and divalent forms of 741F8 anti-c-erbB-2 single-chain FV.", Cancer Res. Sep. 1; 53(17):4026-34 (1993) (Exhibit 1).
Huston, JS et al., "Antigen recognition and targeted delivery by the single-chain Fv"; Cell Biophys. Jan.-Jun.; 22(1-3):189-224 (1993) (Exhibit 2).
Huston, JS, "Multisite association by recombinant proteins can enhance binding selectivity. Preferential removal of immune complexes from serum by immobilized truncated FB analogues of the B domain from staphylococcal protein A", Biophys. J. Apr.;62(1):87-91 (1992) (Exhibit 3).
McCartney, JE et al., "Biosynthetic antibody binding sites; development of a single-chain Fv model based on antidinitrophenol IgA myeloma MOPC 315", J. Protein Chem. Dec;10(6):669-83 (1991) (Exhibit 4).
Tai, MS et al., "A bifunctional fusion protein containing Fc-binding fragment B of staphylococcal protein A amino terminal to antidigoxin single-chain Fv" Biochemistry Sep. 4;29(35):8024-30 (1990) (Exhibit 5).
Huston, JS et al., "Protein engineering of single-chain Fv analogs and fusion proteins", Methods Enzymol, 203:46-88 (1991) (Exhibit 10) Huston, JS et al., "Protein engineering of single-chain Fv analogs and fusion proteins", Methods Enzymol, 203:46-88 (1991) (Exhibit 6).
Anderson et al., "Crosslinking of T3 (CD3) with T4 (CD4) Enhances the Proliferation of Resting T Lymphocytes", J. Immunol. 139:678-682 (1987) (Exhibit 1).
Capon et al., "Designing CD4 Immunoadhesions for AIDS Therapy", Nature 337:525-531 (1989) (Exhbit 2).
Chappel et al., "Identification of the Fc Receptor Class I Binding Site in Human IgG Through the Use of Recombinant IgG/IgG2 Hybrid and Point-Mutated Antibodies", Proc. Natl. Acad. Sci USA 88:9036-9040 (1991) (Exhibit 3).
Davis et al., "Single Chain Antibody (SCA) Encoding Genes: One Step Construction and Expression in Eukaryotic Cells", Biotechnology 9:165-169 (Exhibit 4).
Emmrich et al., "Selective Stimulation of Human T Lymphocyte Subsets by Heteroconjugates of Antibodies to the T-cell Receptor and to Subset-specific Differentiation Antigens", Eur. J. Immunol. 18:645-648 (1988) (Exhibit 5).
Ford et al., "Fusion Tails for the Recovery and Purification of Recombinant Proteins", Protein Expression and Purification 2:95-107 (1991) (Exhibit 6).
Fouser et al., "High Level Expression of a Chimeric Anti-Ganglioside GD2 Antibody: Genomic Kappa Sequences Improve Expression in COS and CHO Cells", Biotechnology 10:1121-1127 (1992) (Exhibit 7).
Hieter et al., "Cloned Human and Mouse Kappa Immunoglobulin Constant and J Region Genes Conserve Homology in Functional Segments" Cell 22:197-207 (1980) (Exhibit 8).
Jung et al., "Induction of Cytotoxicity in Resting Human T Lymphocytes Bound to Tumor Cells by Antibody Heteroconjugates", Proc. Natl. Acad. Sci. USA 84:4611-4615 (1987) (Exhibit 9).
Ledbetter et al., "Signal Transduction Through CD4 Proximity to the CD3/T Cell Receptor", Eur. J. Immunol. 18:525-532 (1988) (Exhibit 10).
Ledbetter et al., "Enhanced Transmembrane Signalling Activity of Monoclonal Antibody Heteroconjugates Suggest Molecular Interactions Between Receptors on the T Cell Surface", Mol. Immunol. 26:137-145 (1989) (Exhibit 11).
Linsley et al., "Binding of the B Cell Activation Antigen B7 to CD28 Costimulates T Cell Proliferation and Interleukin 2 mRNA Accumulation", J. Exp. Med. 173:721-730 (1991) (Exhibit 12).
Liu et al., "Heteroantibody Duplexes Target Cells for Lysis by Cytotoxic T Lymphocytes", Proc. Natl. Acad. Sci. USA 82:8648-8652 (1985) (Exhibit 13).
Liu et al., "Chimeric Mouse-Human IgG1 Antibody that can Mediate Lysis of Cancer Cells", Proc. Natl. Acad. Sci. USA 84:3439-3443 (Exhibit 14).
Lund et al., "Human Fc RI and Fc RII Interact with Distinct but Overlapping Sites on Human IgG", Journal of Immunology 147:2657-2662 (Exhibit 15).
Mayforth et al., "Current Concepts: Designer and Catalytic Antibodies", New Eng. J. Med. 323:173-178 (1990) (Exhibit 16).
Morrison, S. "In Vitro Antibodies: Strategies for Production and Application", Ann. Rev. Immunol. 10:239-266 (1992) (Exhibit 17).
Neuberger et al., "Recombinant Antibodies Possessing Novel Effector Functions", Nature 312:604-608 (1984) (Exhibit 18).
Orlandi et al., "Cloning Immunoglobulin Variable Domains for Expression by the Polymerase Chain Reaction", Proc. Nat. Acad. Sci. 86:3833-3837 (1989) (Exhibit 19).
Perez et al., "Specific Targeting of Cytotoxic T Cells by Anti-T3 Linked to Anti-Target Cell Antibody" Nature 316:354-356 (1985) (Exhibit 20).
Riechmann et al., "Expression of an Antibody Fv Fragment in Myeloma Cells" J. Mol. Biol. 203:825-828 (1988) (Exhibit 21).
Shopes, B. "A Genetically Engineered Human IgG Mutant Cytolytic Activity", Journal of Immunology 9:2918-2922 (1992) (Exhibit 22).
Staerz et al., "Hybrid Hybridoma Producing a Bispecific Monoclonal Antibody that can Focus Effector T-cell Activity", Proc. Natl. Acad. Sci. USA 83:1453-1457 (1986) (Exhibit 23).
Traunecker et al., "Bispecific Single Chain Molecules (Janusins) Target Cytotoxic Lymphocytes on HIV Infected Cells", EMBO 10:3655-3659 (1991) (Exhibit 24).
Emmrich et al., "Synergism in the Activation of Human CD8 T Cells by Cross-Linking the T-Cell Receptor Complex with the CD8 Differentiation Antigen", Proc. Natl. Acad. Sci. USA 83:8298-8302 (1986) (Exhibit 25).
Wels et al., "Construction, Bacterial Expression and Characterization of a Bifunctional Single-Chain Antibody Phosphate Fusion Protein Targeted to the Human ERBB-2 Receptor", Biotechnology 10:1128-1132 (1992) (Exhibit 26).
Winter et al., "Man Made Antibodies", Nature 349:293-299 (1991) (Exhibit 27).
Pluckthun, A. "Mono-and Bivalent Antibody Fragments Produced in Escherichia coli: Engineering, Folding and Antigen Binding", Immunological Reviews 103:151-188 (1992) (Exhibit 28).
Emmrich et al., "Synergism in the Activation of Human CD8 T Cells by Cross-linking the T-Cell Receptor Complex with the CD8 Differentiation Antigen", Proc. Natl. Acad. Sci. USA 83:8298-8302 (1986) (Exhibit 29).
Bird et al., "Single-Chain Antigen-Binding Proteins", Science 24:423-426 (1988) (Exhibit 30).
Hollinger et al., "Diabodies: Small Bivalent and Bispecific Antibody Fragments", Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993) (Exhibit 31).
Huston et al., Methods in Enzymology, vol. 203, 1991, pp. 47-89.

Affiliated with

Also associated with

Rating

Expression vectors encoding bispecific fusion proteins and metho does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Expression vectors encoding bispecific fusion proteins and metho, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Expression vectors encoding bispecific fusion proteins and metho will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-467218