{"search_session":{},"preferences":{"l":"en","queryLanguage":"en"},"patentId":"109-899-059-938-361","frontPageModel":{"patentViewModel":{"ref":{"entityRefId":"109-899-059-938-361","entityRefType":"PATENT"},"entityMetadata":{"linkedIds":{"empty":true},"tags":[],"collections":[{"id":8838,"type":"PATENT","title":"Univ of Illinois Patent Portfolio","description":"","access":"OPEN_ACCESS","displayAvatar":true,"attested":false,"itemCount":10245,"tags":[],"user":{"id":91044780,"username":"Cambialens","firstName":"","lastName":"","created":"2015-05-04T00:55:26.000Z","displayName":"Cambialens","preferences":"{\"usage\":\"public\",\"beta\":false}","accountType":"PERSONAL","isOauthOnly":false},"notes":[{"id":8209,"type":"COLLECTION","user":{"id":91044780,"username":"Cambialens","firstName":"","lastName":"","created":"2015-05-04T00:55:26.000Z","displayName":"Cambialens","preferences":"{\"usage\":\"public\",\"beta\":false}","accountType":"PERSONAL","isOauthOnly":false},"text":"
Search Applicants and Owners separately = \"Univ Illinois\", \"Illinois Univ\", \"University of Illinois\", \"Illinois University\", \"University Illinois NOT Northern NOT southern NOT State\".
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Total patents: 9630
Search Applicants and Owners separately = \"Univ Illinois\", \"Illinois Univ\", \"University of Illinois\", \"Illinois University\", \"University Illinois NOT Northern NOT southern NOT State\".
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Total patents: 9630
expression cassettes capable of being covalently ligated to a gene encoding said wild-type T cell binding protein, each having the structure
5'-GAL 1-10 -- a AGA2p BHA-- mutated polypeptide Bc-myc-3 ;
a vector capable of accepting said expression cassettes;
said vector useable with yeast cells to yield a multiplicity of transformed yeast cells, capable of expressing said cassettes in said transformed yeast cells, whereby said mutated T cell binding proteins are joined at the N-terminus to the C-terminus of a first epitope tag sequence, the N-terminus of said first epitope tag sequence joined to C-terminus of an agglutinin subunit Aga2p sequence, the T cell binding protein sequence being joined at its C-terminus to a second epitope tag, said Aga2p being joined by at least one disulfide bond to an agglutinin subunit Aga1p on said yeast cell surface;
labels for labeling the displayed proteins on said yeast cells by binding a specific label to said displayed proteins, said labels being readable by flow cytometry when used to sort said yeast cells according to their labeling characteristics, said labels cytometrically distinguishable when used to label c-myc and to said T cell binding protein;
instructions for determining the surface expression level of said T cell binding protein in said sorted cells, and for determining the ligand binding characteristics of said T cell expressing an abundance of T cell binding proteins which exhibits enhanced T cell binding characteristics is identified, and for cloning said at least one preferred yeast cell, and for reducing said at least one disulfide bond whereby said fusion protein is released from said yeast cells."],"number":1,"annotation":false,"claim":true,"title":false},{"lines":["A method for using high affinity TCRs to identify ligands comprising:
labeling high affinity TCRs;
contacting said labeled TCRs with ligands;
identifying the ligand with which the labeled TCR is bound."],"number":2,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 2, wherein said label is selected from the group consisting of: fluorescent compounds, chemiluminescent compounds, radioisotopes and chromophores."],"number":3,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 3, wherein said ligands are peptide/MHC ligands."],"number":4,"annotation":false,"claim":true,"title":false},{"lines":["A method of using high affinity TCRs to bind to a selected peptide/MHC ligand comprising:
labeling said high affinity TCRs with a label that binds to the selected peptide/MHC ligand;
contacting said labeled high affinity TCRs with cells containing MHC molecules."],"number":5,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 5, wherein said label is selected from the group consisting of fluorescent compounds, chemiluminescent compounds, radioisotopes and chromophores."],"number":6,"annotation":false,"claim":true,"title":false},{"lines":["The method for blocking autoimmune destruction of cells comprising:
contacting TCRs with high affinity for the site recognized by the T lymphocytes on the surface of a target cell with cells, whereby the autoimmune destruction of cells is blocked."],"number":7,"annotation":false,"claim":true,"title":false},{"lines":["The method for using high affinity TCRs to treat disease comprising:
coupling a TCR having a high affinity for a neoplastic cell surface marker with a therapeutic compound; and
contacting said TCR with cells."],"number":8,"annotation":false,"claim":true,"title":false},{"lines":["Soluble T cell receptors (TCRs) having higher affinity for a ligand than wild type TCRs."],"number":9,"annotation":false,"claim":true,"title":false},{"lines":["The soluble high affinity TCRs of claim 9, wherein said ligand is a peptide/MHC ligand."],"number":10,"annotation":false,"claim":true,"title":false},{"lines":["The soluble high affinity TCRs of claim 9, wherein said high affinity TCR is made by the method comprising: mutagenizing a TCR to create mutant TCR coding sequences; transforming DNA comprising the mutant TCR coding sequences for mutant TCRs into yeast cells; inducing expression of the mutant TCR coding sequences such that the mutant TCRs are displayed on the surface of yeast cells; contacting the yeast cells with a fluorescent label which binds to the peptide/MHC ligand to produce selected yeast cells; and isolating the yeast cells showing the highest fluorescence."],"number":11,"annotation":false,"claim":true,"title":false},{"lines":["The soluble high affinity TCRs of claim 1 isolated by yeast display."],"number":12,"annotation":false,"claim":true,"title":false},{"lines":["A method for cloning the gene for a high affinity TCR mutant into a system that allows expression of the mutant on the surface of T cells comprising:
mutating TCRs to create high affinity TCR mutants;
cloning said TCR mutants into a vector;
transfecting the vector into T cells;
expressing the high affinity TCR mutant on the surface of T cells."],"number":13,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 13, further comprising:
selecting those T cells that are activated by a peptide/MHC ligand more than the wild type."],"number":14,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 14, wherein the transfected/infected T cells are used for recognition of selected peptide-bearing MHC cells."],"number":15,"annotation":false,"claim":true,"title":false},{"lines":["T cells made by the method of claim 13."],"number":16,"annotation":false,"claim":true,"title":false},{"lines":["A DNA sequence encoding a soluble mutant high affinity TCR exhibit higher affinity for its cognate ligand than wild type TCR."],"number":17,"annotation":false,"claim":true,"title":false},{"lines":["The DNA sequence of claim 18 wherein the cognate ligand is a peptide/MHC ligand."],"number":18,"annotation":false,"claim":true,"title":false},{"lines":["The DNA sequence of claim 18 wherein the cognate ligand is superantigen."],"number":19,"annotation":false,"claim":true,"title":false},{"lines":["The DNA sequence of claim 18, which is made by the method comprising:
mutagenizing a TCR to create mutant TCR coding sequences; transforming DNA comprising the mutant TCR coding sequences for mutant TCRs into yeast cells; inducing expression of the mutant TCR coding sequences such that the mutant TCRs are displayed on the surface of yeast cells; contacting the yeast cells with a fluorescent label which binds to the peptide/MHC ligand to produce selected yeast cells; and isolating the yeast cells showing the highest fluorescent and thereafter isolating the mutant TCR coding sequences from the selected yeast cells."],"number":20,"annotation":false,"claim":true,"title":false},{"lines":["A pharmaceutical composition comprising a high affinity TCR in a pharmaceutical carrier."],"number":21,"annotation":false,"claim":true,"title":false},{"lines":["The method of using the pharmaceutical composition of claim 21 comprising administering the composition to a patient."],"number":22,"annotation":false,"claim":true,"title":false},{"lines":["A method of selecting T cell receptors displayed on a yeast cell surface with improved binding properties to desired labels comprising:
transforming yeast cells with a vector which expresses a T cell receptor fused to a yeast cell wall protein, wherein mutagenesis is used to generate a variegated population of mutants of the T cell receptor;
contacting said yeast cells with a label, wherein said label binds to T cell receptors that are displayed on the yeast cell surface and does not bind to T cell receptors that are not displayed on the yeast cell surface;
quantitating said label, wherein a high level of said label indicates the T cell receptor is displayed on a yeast cell surface and has improved binding properties to said label."],"number":23,"annotation":false,"claim":true,"title":false},{"lines":["A DNA library comprising nucleic acids encoding soluble high affinity TCRs, wherein said TCRs are made by the method of mutagenizing a TCR to create mutant TCR coding sequences; transforming DNA comprising the mutant TCR coding sequences for mutant TCRs into yeast cells; inducing expression of the mutant TCR coding sequences such that the mutant TCRs are displayed on the surface of yeast cells; contacting the yeast cells with a fluorescent label which binds to the peptide/MHC ligand to produce selected yeast cells; and isolating the yeast cells showing the highest fluorescence."],"number":24,"annotation":false,"claim":true,"title":false},{"lines":["A method of isolating proteins that bind to a region-specific label comprising:
transforming yeast cells with a vector which expresses a protein to be tested fused to a yeast cell wall protein, wherein mutagenesis is used to generate a variegated population of mutants of the protein to be tested;
contacting said yeast cells with a region-specific label which binds to a specific region of the protein to be tested;
isolating said yeast cells with which said region-specific label is bound."],"number":25,"annotation":false,"claim":true,"title":false},{"lines":["A method for selecting proteins with improved binding to a label, comprising:
transforming yeast cells with a vector which expresses a protein to be tested fused to a yeast cell wall protein, wherein mutagenesis is used to generate a variegated population of mutants of the protein to be tested;
contacting said yeast cells with a label which binds to proteins that are displayed on the yeast cell surface and does not bind to proteins that are not displayed on the yeast cell surface;
isolating said yeast cells with which said label is bound, wherein the presence of said label bound to a protein to be tested indicates said protein to be tested is displayable on the yeast cell surface;
transforming yeast cells with a vector expressing a protein to be tested fused to a yeast cell wall protein, wherein mutagenesis is used to a generate a variegated population of mutants of the protein to be tested;
labeling said yeast cells with a first label, wherein said first label associates with yeast expressing said protein to be tested and does not associate with yeast which do not express said protein to be tested;
isolating said yeast cells with which said first label is associated; and
analyzing and comparing said properties of said mutant protein expressed by yeast with properties of said wild-type protein, wherein yeast cells exhibiting mutant proteins with enhanced properties over the wild-type protein are selected."],"number":26,"annotation":false,"claim":true,"title":false},{"lines":["A library comprising a plurality of different polypeptides displayed on yeast cells."],"number":27,"annotation":false,"claim":true,"title":false},{"lines":["A method of producing a yeast cell displayed variant ligand binding protein with enhanced binding properties relative to a wild-type of said ligand binding protein, the method comprising:
isolating a gene encoding said wild-type binding protein;
creating a library of mutated proteins by randomly mutating said wild-type protein;
incorporating each said mutated protein into respective expression cassettes, each having the structure
5'-GAL 1-10 -- a Aga2p -- mutated polypeptide;
incorporating each said expression cassette into a respective vector;
transforming yeast cells with said cassette-containing vectors to yield a multiplicity of transformed yeast cells;
expressing said cassettes in said transformed yeast cells, whereby said ligand binding protein is displayed on the surface of each said yeast cell, said displayed ligand binding protein containing one of said mutated binding;
labeling the displayed proteins on said yeast cells by binding a specific label to said displayed proteins;
employing flow cytometry to sort said yeast cells according to their labeling characteristics;
determining the surface expression level of said ligand binding protein in said sorted cells;
determining the ligand binding characteristics of said ligand binding protein on the surface of said sorted cells whereby at least one preferred yeast cell expressing an abundance of ligand binding protein which exhibits enhanced ligand binding characteristics is identified; and
cloning said at least one preferred yeast cell."],"number":28,"annotation":false,"claim":true,"title":false},{"lines":["A method of producing a variant T cell binding protein with enhanced T-cell binding properties relative to a wild-type of said T cell binding protein, the method comprising:
isolating a gene encoding said wild-type T cell binding protein;
creating a library of mutated proteins by randomly mutating said wild-type protein;
incorporating each said mutated protein into respective expression cassettes, each having the structure
5'-GAL 1-10 -- a Aga2p --HA-- mutated polypeptide --c-myc-3';
incorporating each said expression cassette into a respective vector; transforming yeast cells with said cassette-containing vectors to yield a multiplicity of transformed yeast cells;
expressing said cassettes in said transformed yeast cells, whereby a unique fusion protein is displayed on the surface of each said yeast cell, said fusion protein containing one of said mutated T cell binding proteins joined at its N-terminus to the C-terminus of a first epitope tag sequence, the N-terminus of said first epitope tag sequence joined to C-terminus of an agglutinin subunit Aga2p sequence, the T cell binding protein sequence being joined at its C-terminus to a second epitope tag, said Aga2p being joined by two disulfide bonds to an agglutinin subunit Aga1p on said yeast cell surface;
labeling the fusion proteins on said yeast cells by binding cytometrically distinguishable labels to said c-myc and to said T cell binding protein;
employing flow cytometry to sort said yeast cells according to their labeling characteristics;
determining the surface expression level of T cell binding protein in said sorted cells; and
determining the ligand binding characteristics of said T cell binding protein on the surface of said sorted cells whereby at least one preferred yeast cell expressing an abundance of fusion protein which exhibits enhanced T cell binding characteristics is identified;
cloning said at least on preferred yeast cell; and
reducing said disulfide bonds whereby said fusion protein is released from said yeast cells."],"number":29,"annotation":false,"claim":true,"title":false},{"lines":["A process of developing a mutant polypeptide exhibiting more favorable binding of a predetermined ligand relative to the binding characteristics of a wild-type of said polypeptide for said ligand, the process comprising:
randomly mutating a predetermined wild-type polypeptide to yield a population of mutated polypeptides;
creating a library of yeast cells, each of which displays on its surface at least one copy of a fusion protein containing one of said mutated polypeptides, the amino acid sequence of said fusion protein consisting of said mutated polypeptide sequence joined at its N-terminus to the C-terminus of an agglutinin subunit Aga2p sequence, said Aga2p being joined by two disulfide bonds to an agglutinin subunit Aga1p on said yeast cell surface, a first epitope tag sequence between said Aga2p and ligand binding polypeptide sequences, and a second epitope tag sequence joined to the C-terminus of said ligand binding polypeptide sequence, wherein a label is bound to at least one of said second epitope tag and said mutant polypeptide;
sorting said yeast cells by flow cytometry;
cloning cells expressing a desired mutant polypeptide;
rescuing and sequencing the DNA sequence coding for said desired mutant polypeptide;
amplifying and expressing said DNA sequence; and
harvesting the desired mutant polypeptide."],"number":30,"annotation":false,"claim":true,"title":false},{"lines":["A gene expression cassette comprising in order:
GAL 1-10 promoter
a Aga2p *
polypeptidewherein \"*\"is nothing or a first epitope tag, and \"‡\" is nothing or a second epitope tag."],"number":31,"annotation":false,"claim":true,"title":false}]}},"filters":{"npl":[],"notNpl":[],"applicant":[],"notApplicant":[],"inventor":[],"notInventor":[],"owner":[],"notOwner":[],"tags":[],"dates":[],"types":[],"notTypes":[],"j":[],"notJ":[],"fj":[],"notFj":[],"classIpcr":[],"notClassIpcr":[],"classNat":[],"notClassNat":[],"classCpc":[],"notClassCpc":[],"so":[],"notSo":[],"sat":[]},"sequenceFilters":{"s":"SEQIDNO","d":"ASCENDING","p":0,"n":10,"sp":[],"si":[],"len":[],"t":[],"loc":[]}}