Protein detection using proximity-dependent DNA ligation assays

• The advent of in vitro DNA amplification has enabled rapid acquisition of genomic information. We present here an analogous technique for protein detection, in which the coordinated and proximal binding of a target protein by two DNA aptamers promotes ligation of oligonucleotides linked to each aptamer affinity probe. The ligation of two such proximity probes gives rise to an amplifiable DNA sequence that reflects the identity and amount of the target protein. This proximity ligation assay detects zeptomole (40 x 10(-21) mol) amounts of the cytokine platelet-derived growth factor (PDGF) without washes or separations, and the mechanism can be generalized to other forms of protein analysis.

Nyckelord

Animals

Base Sequence

Chemistry; Clinical/*methods

DNA/*metabolism

Dose-Response Relationship; Drug

Enzyme-Linked Immunosorbent Assay

Humans

Molecular Sequence Data

Oligonucleotides/*metabolism

Platelet-Derived Growth Factor/*analysis/pharmacology

Proteins/*analysis

Sensitivity and Specificity

Thrombin/pharmacology

Time Factors

Publikations- och innehållstyp

ref (ämneskategori)

art (ämneskategori)