title: Protein detection using proximity-dependent DNA ligation assays.
abstract: 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.
authors: The Beijer Laboratory, Department of Genetics and Pathology, Rudbeck Laboratory, Se-75185 Uppsala, Sweden.
authors: Simon
authors: S
authors: Fredriksson
authors: Mats
authors: M
authors: Gullberg
authors: Jonas
authors: J
authors: Jarvius
authors: Charlotta
authors: C
authors: Olsson
authors: Kristian
authors: K
authors: Pietras
authors: Sigrún Margrét
authors: SM
authors: Gústafsdóttir
authors: Arne
authors: A
authors: Ostman
authors: Ulf
authors: U
authors: Landegren
mesh_terms: D000818:Animals; D001483:Base Sequence; D000077214:Becaplermin; D002624:Chemistry, Clinical / Q000379:methods*; D004247:DNA / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000378:metabolism*; D010982:Platelet-Derived Growth Factor / Q000032:analysis* / Q000494:pharmacology; D011506:Proteins / Q000032:analysis*; D020574:Proto-Oncogene Proteins c-sis; D012680:Sensitivity and Specificity; D013917:Thrombin / Q000494:pharmacology; D013997:Time Factors
chemical_list: D009841:Oligonucleotides; D010982:Platelet-Derived Growth Factor; D011506:Proteins; D020574:Proto-Oncogene Proteins c-sis; D000077214:Becaplermin; D004247:DNA; D013917:Thrombin