Electrochemical biosensor based on enzyme substrate as a linker: Application for aldolase activity with pectin-thionine complex as recognization element and signal amplification probe.
A new strategy to fabricate electrochemical biosensor is reported based on the linkage of enzyme substrate, thereby an electrochemical method to detect aldolase activity is established using pectin-thionine complex (PTC) as recognization element and signal probe. The linkage effect of fructose-1,6-bisphosphate (FBP), the substrate of aldolase, can be achieved via its strong binding to magnetic nanoparticles (MNPs)/aminophenylboronic acid (APBA) and the formation of phosphoramidate bond derived from its reaction with p-phenylenediamine (PDA) on the surface of electrode. Aldolase can reversibly catalyze the substrates into the products which have no binding capacity with MNPs/APBA, resulting in the exposure of the corresponding binding sites and its subsequent recognization on signal probe. Meanwhile, signal amplification can be accomplished by using the firstly prepared PTC which can bind with MNPs/APBA, and accuracy can be strengthened through magnetic separation. With good precision and accuracy, the established sensor may be extended to other proteins with reversible catalyzed ability.
Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, China.
Xiaonan
X
Wang
Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, China; Shanghai Key Laboratory of Bio-Energy Crops, Shanghai University, Shanghai 200444, China.
Meiwen
M
Wang
Department of Obstetrics and Gynecology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210036, China.
Yuanyuan
Y
Zhang
Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, China.
Xiaocao
X
Miao
Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, China.
Yuanyuan
Y
Huang
Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, China. Electronic address: juanzhang@shu.edu.cn.
Juan
J
Zhang
Department of Obstetrics and Gynecology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210036, China. Electronic address: lizhou_sun121@hotmail.com.
Lizhou
L
Sun
D000818:Animals; D001665:Binding Sites; D015374:Biosensing Techniques / Q000379:methods; D001897:Boronic Acids / Q000737:chemistry; D002417:Cattle; D055664:Electrochemical Techniques / Q000379:methods*; D004566:Electrodes; D057075:Enzyme Assays / Q000379:methods; D005634:Fructose-Bisphosphate Aldolase / Q000032:analysis / Q000097:blood* / Q000378:metabolism*; D005635:Fructosediphosphates / Q000737:chemistry / Q000378:metabolism*; D058185:Magnetite Nanoparticles / Q000737:chemistry; D010368:Pectins / Q000737:chemistry*; D010640:Phenothiazines / Q000737:chemistry*; D011817:Rabbits; D013379:Substrate Specificity
D001897:Boronic Acids; D005635:Fructosediphosphates; D058185:Magnetite Nanoparticles; D010640:Phenothiazines; C028592:3-aminobenzeneboronic acid; D010368:Pectins; D005634:Fructose-Bisphosphate Aldolase; C029063:fructose-1,6-diphosphate; C009469:thionine
Aldolase; Electrochemical method; Enzyme substrate linker; Pectin-thionine complex