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Label-free protein recognition using aptamer-based fluorescence assay.
Analyst. 2010 Jul; 135(7):1731-5.A

Abstract

Monitoring proteins in real time and in homogeneous solution without using external labels has always been a difficult task. In this paper, we have developed a label-free method for the ultrasensitive detection of thrombin in homogeneous solutions. High-affinity thrombin-binding aptamer (TBA) used as molecular recognition probe, and fluorophore, crystal violet (CV), was chose as fluorescence signal probe. The fluorescence of CV enhanced significantly when the free CV solution was mixed with single-stranded TBA. In the presence of human thrombin, the fluorescence of CV decreased after the specific interaction between TBA and thrombin. Using the fluorescence change, we are able to selectively detect the thrombin in homogeneous solutions. The conformation transformation was investigated by circular dichroism (CD) spectra measurements. Our method has been shown to be simple and effective without any labelling of the probe or of the target, and this procedure poses minimum effects on the binding properties of the proteins. This assay is highly selective and ultrasensitive. Under the optimum conditions, the method exhibits a dynamic response range from 2 x 10(-11) to 2 x 10(-9) M with a detection limit of 8 x 10(-12) M.

Authors+Show Affiliations

Key Laboratory of Applied Surface and Colloid Chemistry, Ministry of Education, Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Materials Science, Shaanxi Normal University, Xi'an, 710062, China. jinyan@snnu.edu.cnNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20467654

Citation

Jin, Yan, et al. "Label-free Protein Recognition Using Aptamer-based Fluorescence Assay." The Analyst, vol. 135, no. 7, 2010, pp. 1731-5.
Jin Y, Bai J, Li H. Label-free protein recognition using aptamer-based fluorescence assay. Analyst. 2010;135(7):1731-5.
Jin, Y., Bai, J., & Li, H. (2010). Label-free protein recognition using aptamer-based fluorescence assay. The Analyst, 135(7), 1731-5. https://doi.org/10.1039/c0an00014k
Jin Y, Bai J, Li H. Label-free Protein Recognition Using Aptamer-based Fluorescence Assay. Analyst. 2010;135(7):1731-5. PubMed PMID: 20467654.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Label-free protein recognition using aptamer-based fluorescence assay. AU - Jin,Yan, AU - Bai,Jinyan, AU - Li,Hongyan, Y1 - 2010/05/13/ PY - 2010/5/15/entrez PY - 2010/5/15/pubmed PY - 2010/9/30/medline SP - 1731 EP - 5 JF - The Analyst JO - Analyst VL - 135 IS - 7 N2 - Monitoring proteins in real time and in homogeneous solution without using external labels has always been a difficult task. In this paper, we have developed a label-free method for the ultrasensitive detection of thrombin in homogeneous solutions. High-affinity thrombin-binding aptamer (TBA) used as molecular recognition probe, and fluorophore, crystal violet (CV), was chose as fluorescence signal probe. The fluorescence of CV enhanced significantly when the free CV solution was mixed with single-stranded TBA. In the presence of human thrombin, the fluorescence of CV decreased after the specific interaction between TBA and thrombin. Using the fluorescence change, we are able to selectively detect the thrombin in homogeneous solutions. The conformation transformation was investigated by circular dichroism (CD) spectra measurements. Our method has been shown to be simple and effective without any labelling of the probe or of the target, and this procedure poses minimum effects on the binding properties of the proteins. This assay is highly selective and ultrasensitive. Under the optimum conditions, the method exhibits a dynamic response range from 2 x 10(-11) to 2 x 10(-9) M with a detection limit of 8 x 10(-12) M. SN - 1364-5528 UR - https://www.unboundmedicine.com/medline/citation/20467654/Label_free_protein_recognition_using_aptamer_based_fluorescence_assay_ L2 - https://doi.org/10.1039/c0an00014k DB - PRIME DP - Unbound Medicine ER -