Abstract: The present invention provides recombinant peptides that specifically and selectively bind to the human milk fat globule (HMFG) antigen, BA46. In particular, the present invention provides recombinant variants of the Mc3 antibody, including humanized versions of Mc3. The variant Mc3 peptides are particularly useful for diagnostic, prognostic, and therapeutic applications in the field of breast cancer. The present invention also provides methods for the humanization of antibodies such as murine monoclonal antibodies. The novel humanization methods are applied to the production of humanized Mc3 antibodies and it is shown that these humanized antibodies retain the ability to engage in high affinity binding to their cognate antigen. Such humanization enables the use of these antibodies for immunodiagnostic and immunotherapeutic applications in humans.

Abstract: An analogue peptide that comprises the variable regions of the light or heavy chains of an antibody of a first species selectively binding to a carcinoma antigen has 1 to 46 amino acids of the framework regions per chain substituted with amino acids such as those present in equivalent positions in antibodies of a species other than the first species, or fragments thereof comprising 1 to 3 variable region CDRs per chain and optionally flanking regions thereof of 1 to 10 or more amino acids, alone or with an N-terminal fragment of 1 to 10 or more amino acids, combinations or mixtures thereof. The polypeptide may also comprise an effector agent and/or be glycosylated, and is presented as a composition with a carrier. The analogue peptides are used in diagnostic kits for carcinomas and methods for in vivo imaging and treating a primary or metastasized carcinoma, and in vitro diagnosing a carcinoma, ex vivo purging neoplastic cells from a biological fluid.

Abstract: An analogue peptide that comprises the variable regions of the light or heavy chains of an antibody of a first species selectively binding to a carcinoma antigen has 1 to 46 amino acids of the framework regions per chain substituted with amino acids such as those present in equivalent positions in antibodies of a species other than the first species, or fragments thereof comprising 1 to 3 variable region CDRs per chain and optionally flanking regions thereof of 1 to 10 or more amino acids, alone or with an N-terminal fragment of 1 to 10 or more amino acids, combinations or mixtures thereof. The polypeptide may also comprise an effector agent and/or be glycosylated, and is presented as a composition with a carrier. The analogue peptides are used in diagnostic kits for carcinomas and methods for in vivo imaging and treating a primary or metastasized carcinoma, and in vitro diagnosing a carcinoma, ex vivo purging neoplastic cells from a biological fluid.

Abstract: The present invention provides recombinant peptides that specifically and selectively bind to the human milk fat globule (HMFG) antigen, BA46. In particular, the present invention provides recombinant variants of the Mc3 antibody, including humanized versions of Mc3. The variant Mc3 peptides are particularly useful for diagnostic, prognostic, and therapeutic applications in the field of breast cancer.

Abstract: An analogue peptide that comprises the variable regions of the light or heavy chains of an antibody of a first species selectively binding to a carcinoma antigen has 1 to 46 amino acids of the framework regions per chain substituted with amino acids such as those present in equivalent positions in antibodies of a species other than the first species, or fragments thereof comprising 1 to 3 variable region CDRs per chain and optionally flanking regions thereof of 1 to 10 or more amino acids, alone or with an N-terminal fragment of 1 to 10 or more amino acids, combinations or mixtures thereof. The polypeptide may also comprise an effector agent and/or be glycosylated, and is presented as a composition with a carrier. The analogue peptides are used in diagnostic kits for carcinomas and methods for in vivo imaging and treating a primary or metastasized carcinoma, and in vitro diagnosing a carcinoma, ex vivo purging neoplastic cells from a biological fluid.

Abstract: The present invention provides recombinant peptides that specifically and selectively bind to the human milk fat globule (HMFG) antigen, BA46. In particular, the present invention provides recombinant variants of the Mc3 antibody, including humanized versions of Mc3. The variant Mc3 peptides are particularly useful for diagnostic, prognostic, and therapeutic applications in the field of breast cancer. The present invention also provides methods for the humanization of antibodies such as murine monoclonal antibodies. The novel humanization methods are applied to the production of humanized Mc3 antibodies and it is shown that these humanized antibodies retain the ability to engage in high affinity binding to their cognate antigen. Such humanization enables the use of these antibodies for immunodiagnostic and immunotherapeutic applications in humans.

Abstract: A fusion protein comprises a sequence of amino acids which binds antibodies specific to the human milk fat globule (HMFG) differentiation antigens. Specific amino acid sequences are provided, and a fusion protein may be used as an immunogen and for diagnostic purposes.

Abstract: An analogue peptide that comprises the variable regions of the light or heavy chains of an antibody of a first species selectively binding to a carcinoma antigen has 1 to 46 amino acids of the framework regions per chain substituted with amino acids such as those present in equivalent positions in antibodies of a species other than the first species, or fragments thereof comprising 1 to 3 variable region CDRs per chain and optionally flanking regions thereof of 1 to 10 or more amino acids, alone or with an N-terminal fragment of 1 to 10 or more amino acids, combinations or mixtures thereof. The polypeptide may also comprise an effector agent and/or be glycosylated, and is presented as a composition with a carrier. The analogue peptides are used in diagnostic kits for carcinomas and methods for in vivo imaging and treating a primary or metastasized carcinoma, and in vitro diagnosing a carcinoma, ex vivo purging neoplastic cells from a biological fluid.

Abstract: A polynucleotide encodes a modified antibody, or single chains thereof. The modified antibody has a non-antigen-binding peptide such as the constant regions of an antibody of a first species, peptide hormones, enzymes, and peptide transmitters; and a binding peptide such as the unsubstituted light and heavy chains of the variable region of an antibody of a second species which binds the human milk fat globule (HMFG) antigen. The non-antigen-binding peptide is linked to at least one chain of the binding peptide, the chains may be linked to one another at a site other than the antigenic binding site, and at least one chain of the binding peptide has 1 to 46 amino acids substituted with amino acids selected from specific ones assigned to each site. The polynucleotide and other products are also provided in the form of compositions, with a carrier.

Abstract: An improved in vivo method of inhibiting or retarding the growth of malignant tumor cells in a subject comprises administering the subject antitumor therapy, the improvement comprising administering to the subject at least one product comprising an agent selected from the group consisting of polyunsaturated fatty acids, prostaglandins, prostacyclins, thromboxanes, leukotrienes, malone-aldehyde, triene epoxides, analogs thereof, salts thereof and mixtures thereof, the antitumor therapy and the product being administered in amounts and for a period of time effective to attain the desired effect.

Abstract: A polypeptide has the antibody binding activity of the 70K dalton HMFG differentiation antigen and is also provided as a fusion protein with a second antigenic polypeptide. An antibody has affinity for the polypeptide of the invention or a functional fragment thereof. in vivo and in vitro methods for therapy, vaccination and detecting the presence of the polypeptide, the antibody, the DNA and RNA of the invention are provided. DNA and RNA sequences encode the polypeptide of the invention or fragments thereof and immunoassay kits comprise the antibodies and/or polypeptides of the invention.

Abstract: An in vitro solid-phase, competitive assay for detecting the presence of a peptide analyte in a biological sample comprises contacting a fusion protein, of first and second peptides, with a solid supported first antibody or fragment thereof, which binds the first peptide, adding a biological sample containing the peptide analyte and a second antibody or fragment thereof which binds the peptide analyte and the second peptide. The second antibody is allowed to bind any free analyte present in the sample, and to form solid supported complexes of fusion protein and antibody. The amount of solid supported second antibody detected is then compared to a control. When the amount of peptide analyte in the sample increases the amount of second antibody bound to the solid support decreases. An optional final addition of an antibody-binding molecule helps detect the solid supported complexes of the fusion protein and the second antibody or fragment.

Abstract: An in vitro solid-phase, competitive assay for detecting the presence of a peptide analyte in a biological sample, comprising contacting a fusion protein made of a first peptide and a second peptide to a solid supported first antibody which specifically binds to the first peptide, adding thereto a biological sample containing a peptide analyte, adding a second antibody specifically binding to the analyte and the second peptide, and allowing the second antibody to bind any free analyte present in the sample and the solid supported fusion protein to form analyte-second antibody and solid supported fusion protein-second antibody complexes, and determining the amount of solid supported second antibody present and comparing it to a control. When the amount of peptide analyte in the sample increases the amount of a second antibody bound to the solid support decreases. An optional final addition of an antibody-binding molecule helps detect the antibody-fusion protein complex bound to the solid support.

Abstract: A polypeptide has the antibody binding activity of the 46K dalton HMFG differentiation antigen and/or homology to at least one of the light chains of clotting factors V and VIII and is also provided as a fusion protein with a second antigenic polypeptide. An antibody has affinity for the polypeptide of the invention or a functional fragment thereof. in vivo and in vitro methods for therapy vaccination and detecting the presence of the polypeptide, the antibody, the DNA and RNA of the invention are provided. DNA and RNA sequences encode the polypeptide of the invention or fragments thereof and immunoassay kits comprise the antibodies and/or polypeptides of the invention.

Abstract: A hybridoma cell line is provided which is capable of producing monoclonal antibodies which bind to a unique determinant site on the surface and/or in the cytoplasm of human breast carcinoma cells and carcinoma cells of other adenocarcinomas and on the surface of normal human breast epithelial cells. The cell line of the invention was developed by immunizing mice with a select group of immunogens and a conventional myeloma cell line for fusion with the murine splenocytes harvested.The monoclonal antibody is identified as the BrE2 monoclonal antibody. The antigen is characterized as a high molecular weight glycoprotein complex having a molecular exceeding 400,000 daltons. This monoclonal antibody is especially useful for diagnostic, prognostic and therapeutic applications in human breast cell carcinoma.

Abstract: A hybridoma cell line is provided which is capable of producing a monoclonal antibody which binds to a unique determinant site on the surface and in the cytoplasm of human breast carcinoma cells and cells of other adenocarcinomas and does not bind selectively to the surface of normal breast epithelial cells except in instances of high concentration of the antibody in the testing fluid. The cell line of the invention was developed by immunizing mice with a select group of immunogens and a conventional myeloma cell line for fusion with the murine splenocytes harvested. The monoclonal antibody is identified as the BrE3 monoclonal antibody. The BrE3 monoclonal antibody binds to an antigen which is characterized as a high molecular weight glycoprotein complex having a molecular weight exceeding 400,000 daltons that is bound by a disulfide to a protein of 69,000 daltons.

Abstract: Methods and compositions are provided for detecting the presence of carcinomas in a mammalian host by measuring the level of normal surface antigens specific for a differentiated cell in the serum of the host as compared to the normal level of such antigen. The method finds particular use in detecting residual carcinomas after therapy or in detecting the recurrence of neoplastic tissue, and assigning a tissue of origin to the neoplastic tissue.

Abstract: Compositions, articles of manufacture, and methods are provided for enhanced binding of compounds to a surface. The compositions find particular use in coating wells, slides, other surfaces, where ligands or receptors are to be bound and detrimental non-specific binding may be encountered. Particularly, proteins are modified by alkylation to produce a product which strongly adheres to a surface, allows ligands and receptors to be firmly attached by means of the alkylated protein to a surface under mild non-denaturing conditions and permits procedures to minimize non-specific binding of proteins without significant loss of the materials of interest.

Abstract: An anti-KC-4 humanized monoclonal antibody that comprises the variable regions of the light and heavy chains of the anti-KC-4 murine antibody, wherein the light chain has 7 amino acids and the heavy chain has 12 amino acids of the framework regions substituted with amino acid present in equivalent positions in antibodies of a species other than munne, and the constant regions of a human antibody. The antibody may be labeled and/or glycosylated, and is presented as a composition with a carrier. The anti-KC-4 monoclonal antibody is used in diagnostic kits for cancer and in in vivo methods of imaging and treating a primary or metastasized cancer, and in vitro diagnosis and ex vivo purging neoplastic cells from a biological fluid. RNAs and DNAs encode the monoclonal antibody, and a hybrid vector carrying the nucleotides and transfected cells express the peptides.