Signaling aptamer/protein binding by a molecular light switch complex.
A novel method of signaling aptamer/protein binding for aptamer-based protein detection has been developed using a molecular light switch complex, [Ru(phen)2(dppz)]2+. The method takes advantage of the sensitive luminescence signal change of [Ru(phen)2(dppz)]2+ intercalating to the aptamer upon protein/aptamer binding. A 37-nt DNA aptamer against immunoglobulin E (IgE) was first tested as a model system. The luminescence of the [Ru(phen)2(dppz)]2+/IgE aptamer decreased with the increase of IgE. By monitoring the luminescence change, we were able to detect the binding events between the aptamer and IgE for IgE quantitation in homogeneous solutions as well as in serum. The assay was highly selective and sensitive with a detection limit of 100 pM for IgE. This new method is very simple and without the need for the covalent coupling of fluorophores to aptamers. The generalizability of the method was demonstrated by the direct detection of two other proteins, oncoprotein platelet derived growth factor-BB (PDGF-BB) using its DNA aptamer and alpha-thrombin using its RNA aptamer. This new approach is expected to promote the exploitation of aptamer-based biosensors for protein assays in biochemical and biomedical studies.
Jiang
Yaxin
Y
Institute of Chemistry, Chinese Academy of Sciences, Beijing 100080 (China).
Fang
Xiaohong
X
Bai
Chunli
C
D052157:Aptamers, Nucleotide / Q000032:analysis* / Q000737:chemistry; D000077214:Becaplermin; D002021:Buffers; D004247:DNA / Q000032:analysis / Q000737:chemistry; D006801:Humans; D007073:Immunoglobulin E / Q000032:analysis / Q000737:chemistry; D008027:Light; D008163:Luminescent Measurements / Q000379:methods*; D009942:Organometallic Compounds / Q000737:chemistry*; D009994:Osmolar Concentration; D010982:Platelet-Derived Growth Factor / Q000032:analysis / Q000737:chemistry; D011485:Protein Binding; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D020574:Proto-Oncogene Proteins c-sis; D011522:Protons; D012313:RNA / Q000032:analysis / Q000737:chemistry; D012680:Sensitivity and Specificity; D013917:Thrombin / Q000032:analysis / Q000737:chemistry
D052157:Aptamers, Nucleotide; D002021:Buffers; D009942:Organometallic Compounds; D010982:Platelet-Derived Growth Factor; D011506:Proteins; D020574:Proto-Oncogene Proteins c-sis; D011522:Protons; C466600:Ru(phen)(2)dipyrido(3,2-a-2',3'-c)phenazine; D000077214:Becaplermin; D007073:Immunoglobulin E; D012313:RNA; D004247:DNA; D013917:Thrombin