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.
Fredriksson
Simon
S
The Beijer Laboratory, Department of Genetics and Pathology, Rudbeck Laboratory, Se-75185 Uppsala, Sweden.
Gullberg
Mats
M
Jarvius
Jonas
J
Olsson
Charlotta
C
Pietras
Kristian
K
Gústafsdóttir
Sigrún Margrét
SM
Ostman
Arne
A
Landegren
Ulf
U
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
D009841:Oligonucleotides; D010982:Platelet-Derived Growth Factor; D011506:Proteins; D020574:Proto-Oncogene Proteins c-sis; D000077214:Becaplermin; D004247:DNA; D013917:Thrombin