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5045451 Methods for screening antibodies for use as immunotoxins
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PATENTABSTRACTS single base change and on the ability to detect the separate mutant molecules. The present invention, in another aspect, relates to a method for determining a mutational spectrum in a DNA sequence of interest present in a population of cells. The method of the present invention is useful as a diagnostic or analytical tool in forensic science in assessing environmental and/or occupational exposures to potentially genetically toxic materials (also referred to as potential mutagens); in biotechnology, particularly in the study of the relationship between the amino acid sequence of enzymes and other biologicallyactive proteins or protein-containing substances and their respective functions; and in determining the effects of drugs, cosmetics and other chemicals for which toxicity data must be obtained.
The invention is directed to hydridoma cell lines and the monoclonal antibodies produced by the cell lines, where the monoclonal antibodies specifically bind to the lipopolysaccharidc cnd~, toxin binding receptor of mammalian cells Specifically, the monoclonal antibodies binds to a cell receptor which has a molecular mass ot 8(t kilodaltons, and is specific ['or the lipid A component of the lipopolysaccharideendotoxin The monoclonal antibodies arc specific ft)r a carbohydrate component of the binding receptor in one instance, and for a protein/peptide receptor in another.
5045451
5045467
METHODS FOR SCREENING ANTIBODIES FOR USE AS IMMUNOTOXINS
SERUM-FREE GROWTH MEDIUM A N D U S E THEREOF
Jonathan W Uhr, Ellen S Vitetta assigned to Board of Regents The present disclosure described an assay for screening monoclonal antibodies for their potential as highly cytotoxic immunotoxins. The assay involves treating cells with dilutions of the test antibody followed by a Fab fragment of a secondary antibody coupled to an A chain toxin ( indirect assay ), The cytotoxicity of the indirect assay is compared to that of the direct assay where the monoclonal antibody is coupled to an A chain toxin. Indirect and direct assays were carried out using 14 antibodies and a panel of 8 human and mouse cell types. The two assays showed v rtually 1000; correlat on The indirect assay, therefore, predicts the potency of a given monoclonal antibody to make an effective immunotoxin and should be useful in screening monoclonal antibodies for use as immunotoxins.
5045466 PURIFIED MAMMALIAN C E L L BINDING RECEPTOR FOR BACTERIAL LIPOPOLYSACCHARIDE ENDOTOXIN A N D
MONOCLONAL ANTIBODY David Morrison, Taiying Chen, Mei-Guey Lei, Stuart Bright, Linda Flebbe assigned to University of Kansas
Kjel Bertheussen, Eidkjosen. Norway assigned to Medi-Cult A/S; A/S GEA Farmaceulisk Fabr Serum-free growth medium comprising an ironchelate, aurin-tricarboxylic acid and optionally alkali-metal-EDTA and trace elements together with possible growth factors, wherein the ironchelate may comprise a mixture of Fe-EDTA and citric acid. The growth medium may be used for quality testing of other growth media, optionally together with a hybridoma cell line from p3U1 tumor cells and B-lymphocytes. which hybridoma also is disclosed.
5045468 PROTEIN-FREE CULTURE MEDIUM WHICH PROMOTES HYBRIDOMA GROWTH Frederick J Darfler assigned to Cell Enterprises Inc A chemically-defined, stable, protein-free medium has been devised which is capable of supporting the growth of lymphoid cells, particularly hybridomas, even in anchorageindependent cultures. Doubling rates are close to those obtained with serum-supplemented media. The medium preferably comprises a nonproteinaceous organo-iron compound, espe-
PATENTABSTRACTS single base change and on the ability to detect the separate mutant molecules. The present invention, in another aspect, relates to a method for determining a mutational spectrum in a DNA sequence of interest present in a population of cells. The method of the present invention is useful as a diagnostic or analytical tool in forensic science in assessing environmental and/or occupational exposures to potentially genetically toxic materials (also referred to as potential mutagens); in biotechnology, particularly in the study of the relationship between the amino acid sequence of enzymes and other biologicallyactive proteins or protein-containing substances and their respective functions; and in determining the effects of drugs, cosmetics and other chemicals for which toxicity data must be obtained.
The invention is directed to hydridoma cell lines and the monoclonal antibodies produced by the cell lines, where the monoclonal antibodies specifically bind to the lipopolysaccharidc cnd~, toxin binding receptor of mammalian cells Specifically, the monoclonal antibodies binds to a cell receptor which has a molecular mass ot 8(t kilodaltons, and is specific ['or the lipid A component of the lipopolysaccharideendotoxin The monoclonal antibodies arc specific ft)r a carbohydrate component of the binding receptor in one instance, and for a protein/peptide receptor in another.
5045451
5045467
METHODS FOR SCREENING ANTIBODIES FOR USE AS IMMUNOTOXINS
SERUM-FREE GROWTH MEDIUM A N D U S E THEREOF
Jonathan W Uhr, Ellen S Vitetta assigned to Board of Regents The present disclosure described an assay for screening monoclonal antibodies for their potential as highly cytotoxic immunotoxins. The assay involves treating cells with dilutions of the test antibody followed by a Fab fragment of a secondary antibody coupled to an A chain toxin ( indirect assay ), The cytotoxicity of the indirect assay is compared to that of the direct assay where the monoclonal antibody is coupled to an A chain toxin. Indirect and direct assays were carried out using 14 antibodies and a panel of 8 human and mouse cell types. The two assays showed v rtually 1000; correlat on The indirect assay, therefore, predicts the potency of a given monoclonal antibody to make an effective immunotoxin and should be useful in screening monoclonal antibodies for use as immunotoxins.
5045466 PURIFIED MAMMALIAN C E L L BINDING RECEPTOR FOR BACTERIAL LIPOPOLYSACCHARIDE ENDOTOXIN A N D
MONOCLONAL ANTIBODY David Morrison, Taiying Chen, Mei-Guey Lei, Stuart Bright, Linda Flebbe assigned to University of Kansas
Kjel Bertheussen, Eidkjosen. Norway assigned to Medi-Cult A/S; A/S GEA Farmaceulisk Fabr Serum-free growth medium comprising an ironchelate, aurin-tricarboxylic acid and optionally alkali-metal-EDTA and trace elements together with possible growth factors, wherein the ironchelate may comprise a mixture of Fe-EDTA and citric acid. The growth medium may be used for quality testing of other growth media, optionally together with a hybridoma cell line from p3U1 tumor cells and B-lymphocytes. which hybridoma also is disclosed.
5045468 PROTEIN-FREE CULTURE MEDIUM WHICH PROMOTES HYBRIDOMA GROWTH Frederick J Darfler assigned to Cell Enterprises Inc A chemically-defined, stable, protein-free medium has been devised which is capable of supporting the growth of lymphoid cells, particularly hybridomas, even in anchorageindependent cultures. Doubling rates are close to those obtained with serum-supplemented media. The medium preferably comprises a nonproteinaceous organo-iron compound, espe-