Differentiation and detection of PDGF isomers and their receptors by tunable aptamer capillary electrophoresis.
Tunable aptamer capillary electrophoresis (CE) techniques were developed to enable the separation and detection of platelet derived growth factor (PDGF) isomers and their receptors. Using an aptamer that formed a stable complex with the B chain but not with the A chain of PDGF, we were able to tweak the electrophoretic mobilities of the PDGF isomers for their separation. PDGF-AB bound to a single aptamer molecule was well resolved from PDGF-BB bound to two aptamer molecules. Simultaneous determination of 50 pM of two isomers was accomplished in a single analysis. Furthermore, PDGF-AB was used as a connector to bring receptor alpha and fluorescent aptamer into a single complex molecule. As a result, the formation of a (receptor alpha)-(PDGF-AB)-(fluorescent aptamer) ternary complex enabled the detection of the receptor alpha by tunable aptamer CE. A competitive assay was developed to determine receptor beta, making use of the competition between the receptor beta and fluorescent aptamer in binding to PDGF-BB. Detection limits were 0.5 nM for PDGF receptor alpha and 3 nM for receptor beta. Determination of PDGF isomers and their receptors in diluted serum samples showed no interference from the sample matrix.
Zhang
Hongquan
H
Division of Analytical and Environmental Toxicology, Department of Laboratory Medicine and Pathology, Faculty of Medicine and Dentistry, 10-102 Clinical Sciences Building, University of Alberta, Edmonton, Alberta, Canada T6G 2G3.
Li
Xing-Fang
XF
Le
X Chris
XC
D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000378:metabolism*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D010982:Platelet-Derived Growth Factor / Q000032:analysis* / Q000737:chemistry; D020033:Protein Isoforms / Q000032:analysis / Q000737:chemistry; D017479:Receptors, Platelet-Derived Growth Factor / Q000032:analysis* / Q000097:blood / Q000737:chemistry; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry
D052157:Aptamers, Nucleotide; D005456:Fluorescent Dyes; D010982:Platelet-Derived Growth Factor; D020033:Protein Isoforms; D011994:Recombinant Proteins; D017479:Receptors, Platelet-Derived Growth Factor