Selection Of DNA Aptamers Against UPAR With Specificity

Posted on:2015-05-09

Degree:Master

Type:Thesis

Country:China

Candidate:H T Yang

Full Text:PDF

GTID:2284330422487786

Subject:Surgery

Abstract/Summary:

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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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