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Selection Of DNA Aptamers Against UPAR With Specificity

Posted on:2015-05-09Degree:MasterType:Thesis
Country:ChinaCandidate:H T YangFull Text:PDF
GTID:2284330422487786Subject:Surgery
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ObjectiveToselectaptamersthatcanbeabletobindtourokinase‐typeplasminogenactivatorreceptor(uPAR)withSpecificitybyusingbead‐basedSystematicEvolutionofLigandsbyExponentialenrichment (SELEX).MethodsInitialDNAlibrarywith87bplengthwaschemicallysynthesized.uPAR‐beadthatrecombinantbyuPARproteinandNi‐beadisusedasthetargetwhileNi‐beadasthecountertarget.Severalotherproteins(EGFR‐bead, GPC3‐beadandThrombin‐bead) anduPAR‐positive(U87, U251, SW480andA549)wereusedtoidentifytheSpecificityoftheaptamerdevelopedinthiswork.Flowcytometricanalysiswasusedtomonitortheenrichmentofaptamersduringoftheselectionandthespecificityoftheaptamerdevelopedinthisselection.ResultswehadidentifiedanaptamernamedDZ‐u1thatspecificallyrecognizedtheproteinofuPAR‐beadandtheuPARpositivecancercelllinesbuthadn’totherproteins.ConclusionApamerDZ‐u1couldspecificallybindtotargetproteinuPARanduPARpositivecelllines.Theaptamerhadthepotentialtobeusedintumorearlydiagnosisandtargettherapy.
Keywords/Search Tags:uPAR, SELEX, Aptamer
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