title: Chemiluminescence detection of protein in capillary electrophoresis using aptamer-functionalized gold nanoparticles as biosensing platform.
abstract: Highly sensitive and selective detection of disease-related proteins play critical roles in clinical practice and diagnostic assays. Herein, we proposed a highly selective and ultrasensitive chemiluminescence (CL) method for protein detection in capillary electrophoresis (CE) using aptamer-functionalized gold nanoparticles (AuNPs) as biosensing platform. In this protocol, AuNPs were synthesized and conjugated with aptamer to form AuNPs-aptamer. Using thrombin and thrombin binding aptamer as an initial proof-of-concept recognization pair, AuNPs-aptamer was linked to thrombin to produce an AuNPs-aptamer-thrombin complex. The resulted complex and unbound AuNPs-aptamer were separated in CE and detected with luminol-H2O2 CL system. The developed strategy produced an ultrasensitive detection of thrombin down to 13.5 fmol/L (S/N=3) with a linear range from 0.033 to 66.0 pmol/L. The application of the present protocol was demonstrated by analyzing thrombin in human plasma samples with the recoveries of 87.6-116.8%. This novel strategy has many outstanding merits including high specificity of aptamer, excellent catalysis behavior of AuNPs, high sensitivity of CL detection, and high separation efficiency of CE.
authors.affiliation: College of Chemistry and Chemical Engineering, Xinyang Normal University, Xinyang 464000, China. Electronic address: liuym9518@sina.com.
authors.forename: Yanming
authors.initials: Y
authors.lastname: Liu
authors.affiliation: College of Chemistry and Chemical Engineering, Xinyang Normal University, Xinyang 464000, China.
authors.forename: Yingying
authors.initials: Y
authors.lastname: Liu
authors.affiliation: College of Chemistry and Chemical Engineering, Xinyang Normal University, Xinyang 464000, China.
authors.forename: Min
authors.initials: M
authors.lastname: Zhou
authors.affiliation: College of Chemistry and Chemical Engineering, Xinyang Normal University, Xinyang 464000, China.
authors.forename: Kejing
authors.initials: K
authors.lastname: Huang
authors.affiliation: College of Chemistry and Chemical Engineering, Xinyang Normal University, Xinyang 464000, China.
authors.forename: Juntao
authors.initials: J
authors.lastname: Cao
authors.affiliation: College of Chemistry and Chemical Engineering, Xinyang Normal University, Xinyang 464000, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.affiliation: Xinyang Central Hospital, Xinyang 464000, China.
authors.forename: Yonghong
authors.initials: Y
authors.lastname: Chen
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry; D015374:Biosensing Techniques / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods*; D006046:Gold / Q000737:chemistry; D006801:Humans; D006861:Hydrogen Peroxide; D008163:Luminescent Measurements / Q000379:methods*; D008165:Luminol; D053768:Metal Nanoparticles / Q000737:chemistry / Q000648:ultrastructure; D013917:Thrombin / Q000032:analysis* / Q000737:chemistry
chemical_list: D052157:Aptamers, Nucleotide; C081676:thrombin aptamer; D008165:Luminol; D006046:Gold; D006861:Hydrogen Peroxide; D013917:Thrombin
keywords: Aptamer-functionalized gold nanoparticles; Capillary electrophoresis; Chemiluminescence; Human plasma determination