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Anti-AChE interference method capable of quickly detecting BChE activity

An active and rapid technology, applied in the field of enzyme activity detection, to achieve the effect of easy operation, good biocompatibility and simple process

Inactive Publication Date: 2013-05-08
NANKAI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, most of the existing methods for detecting cholinesterase activity are aimed at the detection of AChE activity, and there is currently no method for the rapid and direct selective detection of BChE

Method used

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  • Anti-AChE interference method capable of quickly detecting BChE activity
  • Anti-AChE interference method capable of quickly detecting BChE activity
  • Anti-AChE interference method capable of quickly detecting BChE activity

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Embodiment

[0020] A method that is resistant to AChE (acetylcholinesterase) interference and can quickly detect BChE (butyrylcholinesterase) activity, using sulfonated calix[4]arene (SC4A) and dye lucigenin (LCG) as the enzyme activity detection The sensor pair uses succinylcholine (SuCh) as the substrate for the enzyme reaction to establish a supramolecular tandem detection system for BChE activity monitoring. The steps are as follows:

[0021] 1) At 37°C, use 10mmol / L pH8.0 phosphate buffer solution to configure a solution containing 0.5μmol / LSC4A, 0.5μmol / L LCG and 10μmol / L SuCh to obtain a supramolecular tandem for BChE activity monitoring Detection system; the configuration method of the 10mmol / L pH8.0 phosphate buffer solution is: weigh 1.56g sodium dihydrogen phosphate and 0.33g sodium hydroxide to prepare 1L10mmol / L pH8.0 phosphate buffer solution; choose SC4A and LCG as For the sensing pair in the detection process, SC4A and LCG, SuCh, Ch have large bonding constants (K S For 10 5 ...

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Abstract

The invention discloses an anti-AChE interference method capable of quickly detecting BChE activity. The method comprises the following steps of: 1) with sulfonated calix[4]arene and dye lucigenin as a sensing pair for enzyme activity detection and succinylcholine as an enzyme reaction substrate, preparing a solution containing 0.5(mu)mol / L SC4A, 0.5(mu)mol / L LCG and 10(mu)mol / L SuCh by use of 10mmol / L phosphoric acid buffer solution with pH being 8.0 at 37 DEG C, and establishing a supermolecule serial detection system; and 2) adding 1U / ml BChE into the system to obtain an obvious fluorescence quenching signal within 15 minutes. The method disclosed by the invention has the advantages of simple technology, convenience in operation and sensitivity and high speed, can detect the activity of BChE with high selectivity in the presence of excessive AChE, and has broad application prospect in the specific detection of BChE activity and treatment field of Alzheimer's disease.

Description

【Technical Field】 [0001] The invention belongs to enzyme activity detection technology, in particular to a method that is resistant to AChE (acetylcholinesterase) interference and can quickly detect BChE (butyrylcholinesterase) activity. 【Background technique】 [0002] In recent years, due to the importance of cholinesterase in neurotransmitter sensing in the human body and its key role in the treatment of Alzheimer's disease, it has received extensive attention. See: Lane, RM; Potkin, SG; Enz, A .Int.J.Neuropsychopharmacol.2006,9,101–124. At present, the detection based on cholinesterase activity and the screening of corresponding inhibitors have become research hotspots, see: 1) Pardo-Yissar, V.; Katz, E.; Wasserman, J.; Willner, IJAm.Chem. Soc. 2003, 125, 622-623; 2) Maeda, H.; Mastuno, H.; Ushida, M.; Katayama, K.; Saeki, K.; Itoh, N. Angew. Chem., Int. Ed. 2005, 44,2922–2925; 3) Bailey, DM; Hennig, A.; Uzunova, VD; Nau, WMChem. Eur. J. 2008, 14, 6069–6077; 4) Nau, WM; Ghal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 刘育郭东升杨婧
Owner NANKAI UNIV
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