Cancer Research Fund of Contra Costa

Other

Johns Hopkins University

Cancer Research Institute of Contra Costa

John Muir Cancer & Aging Institute

Senomed, Inc.

Cincinnati Children's Hospital Medical Center

Cancer Research Fund

Jerry A Peterson

Roberto L Ceriani

David J Larocca

Fernando Jose Rebelo Do Couto

David S Newburg

Robert H Yolken

Eduardo Agustin Padlan

Robert L Ceriani

Eduardo A Padlan

Robeto L Ceriani

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.

Recombinant peptides derived from the Mc3 anti-BA46 antibody, methods of use thereof, and methods of humanizing antibody peptides

A polypeptide has the antibody binding activity of the 46 Kdalton 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.

Antibody with 46 kdalton HMFG antigen binding specificity, immunoassay kit and diagnostic method

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. RNAs and DNAs encode the analogue peptide, and a hybrid vector carrying the nucleotides and transfected cells express the peptides and a method produces the analogue peptide. An anti-idiotype polypeptide comprises polyclonal antibodies raised against an anti-carcinoma antibody or the analogue peptide of this invention, monoclonal antibodies thereof, Fab, Fab', (Fab&#x2032;), CDR, variable region, or analogues or fragments thereof, combinations thereof with an oligopeptide comprising a TRP trimer, tandem repeats thereof, or combination or mixtures thereof. An anti-idiotype hybrid polypeptide with an effector agent and the anti-idiotype polypeptide, an anti-carcinoma vaccine, an anti-carcinoma vaccination kit, a method of vaccinating against carcinoma and a method of lowering the serum concentration of a circulating antibody or polypeptide are provided.

Modified antibodies with human milk fat globule specificity and uses

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. RNAs and DNAs encode the analogue peptide, and a hybrid vector carrying the nucleotides and transfected cells express the peptides and a method produces the analogue peptide. An anti-idiotype polypeptide comprises polyclonal antibodies raised against an anti-carcinoma antibody or the analogue peptide of this invention, monoclonal antibodies thereof, Fab, Fab&prime;, (Fab&prime;),, CDR, variable region, or analogues or fragments thereof, combinations thereof with an oligopeptide comprising a TRP trimer, tandem repeats thereof, or combination or mixtures thereof. An anti-idiotype hybrid polypeptide with an effector agent and the anti-idiotype polypeptide, an anti-carcinoma vaccine, an anti-carcinoma vaccination kit, a method of vaccinating against carcinoma and a method of lowering the serum concentration of a circulating antibody or polypeptide are provided.

Use of modified antibodies with human milk fat globule specificity

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.

Fusion protein containing HMFG Epitop E (S)

A polypeptide has the antibody binding activity of the 46 Kdalton HMFG antigen and/or homology to at least one of the light chains of clotting factors V and VIII and/or contains RGD and/or EGF-like segments. The polypeptide is provided as a recombinant and/or glycosylated and/or fusion protein. An antibody has high affinity for specificity epitopes of the polypeptide of the invention. Polynucleotide segments encode the polypeptide, recombinant and fusion protein of the invention or fragments thereof, and immunoassay kits comprise the antibodies and/or polypeptides of the invention and other components. In vivo, ex vivo, and in vitro methods of therapy, vaccination and diagnosis utilize the polypeptide, fusion protein anti-sense nucleotides, antibodies or and polynucleotides of the invention.

46 kilodalton human milk fat globule (HMFG) antigen, fragments and

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. RNAs and DNAs encode the analogue peptide, and a hybrid vector carrying the nucleotides and transfected cells express the peptides and a method produces the analogue peptide. An anti-idiotype polypeptide comprises polyclonal antibodies raised against an anti-carcinoma antibody or the analogue peptide of this invention, monoclonal antibodies thereof, Fab, Fab', (Fab').sub.2, CDR, variable region, or analogues or fragments thereof, combinations thereof with an oligopeptide comprising a TRP trimer, tandem repeats thereof, or combination or mixtures thereof. An anti-idiotype hybrid polypeptide with an effector agent and the anti-idiotype polypeptide, an anti-carcinoma vaccine, an anti-carcinoma vaccination kit, a method of vaccinating against carcinoma and a method of lowering the serum concentration of a circulating antibody or polypeptide are provided.

Modified antibodies with human milk fat globule specificity

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. The polynucleotides may be RNAs and DNAs, and are also provided as hybrid vectors carrying them, and as transfected cells expressing the modified antibodies or their single chains.

Polynucleotides encoding modified antibodies with human milk fat globule

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. An in vivo method of inhibiting or retarding the growth of malignant tumor cells comprises administering to a subject in need of such treatment at least one anti-cell antigen specific radiolabeled antibody and at least one product comprising an agent such as polyunsaturated fatty acids, prostaglandins, prostacyclins, thromboxanes, leukotrienes, malonealdehyde, triene epoxides, salts thereof and mixtures thereof, the antibody and the product being administered in amount and for a period of time effective to attain the desired effect. A kit for use in antitumor therapy comprises in separate containers an antitumor agent such as a chemotherapeutic agent, an antitumor hormonal agent, an anti-target cell antibody, external and an internal irradiation agents, or biological response modifiers, and an agent such as polyunsaturated fatty acids, prostaglandins, prostacyclins, thromboxanes, leukotrienes, malonealdehyde, triene epoxides, salts thereof or mixtures thereof.

Method of treating human breast cancer by administration of radiolabeled

An anti-diarrheic product comprises a foodstuff and an anti-rotaviral agent such as human defatted fat globule membranes, the human milk macromolecular fraction, skim milk, curd, whey, the milk mucin complex, the 46 Kd app.MW glycoprotein, a polypeptide having an amino acid sequence having the rotavirus binding specificity of the 46 Kd app.MW HMFG glycoprotein or mixtures thereof. An anti-diarrheal kit comprises the anti-rotaviral agent of this invention and instructions for its use, and optionally a foodstuff. A method for inhibiting or countering rotavirus infection in mammalian cells comprises contacting the cells with an anti-rotaviral amount of the agent of this invention. A therapeutic method for inhibiting the onset of or countering rotavirus infection comprises administering to a subject afflicted with or at risk for rotavirus infections such as those occuring, for example, in infants and children (infantile gasteroenteritis) and immunodeficient persons or those who have received a transplant, an anti-rotaviral effective amount of the agent of this invention.

Anti-viral composition and kit and use for treating rotavirus infection

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.

Nucleic acids and vector encoding HMFQ polypeptide, transfected host,

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. The assay may be utilized for detecting neoplastic antigens from tumors or their metastases by using a second peptide that binds specifically to an antibody specific to an antigen, such as an epithelial mammary cell antigen. A kit for detecting the presence of neoplastic tissue comprises the fusion protein, a first antibody or fragment thereof specifically binding the first peptide, and a second antibody or fragment thereof specifically binding the second peptide, and optionally a device for detecting the second antibody or fragment thereof, such as a labeled antibody-binding molecule.

Solid-phase competitive assay utilizing a fusion protein

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. The assay may be utilized for detecting neoplastic antigens from tumors or their metastases by utilizing a fusion protein containing an antigen of an epithelial mammary cell. A kit for detecting the presence of a neoplastic tissue comprises the fusion protein along with a first antibody specifically binding to the first peptide, and a second antibody specifically binding to the second peptide, and optionally a means for detecting the second antibody or fragment thereof, such as an antibody-binding molecule.

Highly sensitive and specific solid-phase competitive assay utilizing a

An anti-diarrheic product comprises a foodstuff and an anti-rotaviral agent such as human defatted fat globule membranes, the human milk macromolecular fraction, the milk mucin complex, the 46 Kd apparent MW glycoprotein, a polypeptide having the rotavirus-binding specificity of the 46 Kd apparent MW HMFG glycoprotein, mixtures thereof, or mixtures thereof, and optionally skim milk, curd, and/or whey. They product of the invention is provided also as an anti-diarrheal kit, with instructions for its use. The product of the invention has therapeutic and prophylactic application for inhibiting the onset of, or countering, rotavirus infection and/or diarrhea, in a subject, such as for example, infants and children (infantile gasteroenteritis), travellers, and immunodeficient persons, including HIV positive and transplant patients.

Growing community of inventors

Lafayette, CA, United States of America

Jerry A Peterson