title: Simple and sensitive electrogenerated chemiluminescence peptide-based biosensor for detection of matrix metalloproteinase 2 released from living cells.
abstract: A simple and sensitive electrogenerated chemiluminescence biosensor was developed to monitor matrix metalloproteinase 2 (MMP-2) by employing a specific peptide (CGPLGVRGK) as a molecular recognition substrate. Bis(2,2'-bipyridine)-4'-methyl-4-carboxybipyridine-ruthenium N-succinimidyl ester-bis(hexafluorophosphate) (Ru(bpy)2(mcbpy-O-Su-ester)(PF6)2 (Ru1) was used as ECL-emitting species and covalently labeled onto the peptide through NH2-containing lysine on the peptide via acylation reaction to form Ru1-peptide as an ECL probe. An ECL peptide-based biosensor was fabricated by self-assembling the ECL probe onto the surface of gold electrode. MMP-2 can specifically cleave the Ru1-peptide on the electrode surface, which led the partly Ru1-peptide to leave the electrode surface and resulted in the decrease of the ECL intensity obtained from the resulted electrode in 0.1 M phosphate-buffered saline (pH 7.4) containing tri-n-propylamine. The decreased ECL intensity was piecewise linear to the concentration of MMP-2 in the range from 1 to 500 ng/mL. Moreover, the ECL biosensor is successfully applied to detection of MMP-2 secreted by living cell, such as HeLa cells. Additionally, the biosensor was also applied to the evaluation of matrix metalloproteinase inhibitors. The strategy presented here is promising for other disease-related matrix metalloproteinase assay and matrix metalloproteinase inhibitor profiling with sensitivity and simplicity. Graphical Abstract Detection of MMP-2 released from living cells by ECL peptide-based biosensor.
authors.affiliation: Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an, 710062, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Dang
authors.affiliation: Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an, 710062, China.
authors.forename: Hongfang
authors.initials: H
authors.lastname: Gao
authors.affiliation: Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an, 710062, China.|College of Chemical Engineering and Modern Materials, Shangluo University, Shangluo, 726000, China.
authors.forename: Zhejian
authors.initials: Z
authors.lastname: Li
authors.affiliation: Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an, 710062, China. honglanqi@snnu.edu.cn.
authors.forename: Honglan
authors.initials: H
authors.lastname: Qi
authors.affiliation: Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an, 710062, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Gao
authors.affiliation: Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an, 710062, China.
authors.forename: Chengxiao
authors.initials: C
authors.lastname: Zhang
mesh_terms: D015082:2,2'-Dipyridyl / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D015374:Biosensing Techniques / Q000379:methods*; D004566:Electrodes; D057075:Enzyme Assays / Q000379:methods; D006046:Gold / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D049408:Luminescent Agents / Q000737:chemistry* / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods*; D020778:Matrix Metalloproteinase 2 / Q000032:analysis* / Q000378:metabolism; D009942:Organometallic Compounds / Q000737:chemistry / Q000378:metabolism; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D049408:Luminescent Agents; D009942:Organometallic Compounds; D010455:Peptides; C103599:bis(bipyridyl)ruthenium(II); D015082:2,2'-Dipyridyl; D006046:Gold; D020778:Matrix Metalloproteinase 2
keywords: Biosensor; Electrogenerated͘͘͘ chemiluminescence; Matrix metalloproteinase 2; Peptide