Presentation Title

Thread-Based Microfluidic Chips as a Platform to Assess Acetylcholinesterase Activity

Start Date

November 2016

End Date

November 2016

Location

Watkins 2141

Type of Presentation

Oral Talk

Abstract

The quantitative detection of acetylcholinesterase (AChE) by colorimetric analysis utilizing a microfluidic thread-based analytical device (µTAD) is described. It can be difficult to study neurotransmitters (e.g. acetylcholine [ACh]) directly, and therefore the activity of AChE can be studied. Levels of ACh are relevant to Alzheimer’s disease as levels can drop by as much as 90% in patients. One way to compensate for lower levels of ACh is to inhibit the AChE enzyme from breaking down ACh. Hence, the development of facile methods to detect AChE and screen potential inhibitors will aid in the diagnosis (and eventual cure) for Alzheimer’s disease. In the present work, two µTAD platforms were fabricated. Both had a nylon thread trifurcated into three channels terminating at open sites. One platform had one end of the thread split into two (third end cut off). The nylon on the second platform was not trifurcated at the other end, creating a single inlet. 5,5’-Dithiobis-(2-nitrobenzoic acid) (DTNB) was spotted on the analysis sites. Acetylthiocholine iodide (ATC) (or cysteine, Cys), is transported through one channel of the nylon thread by capillary action due to the hydrophilic nature of nylon. AChE is transported through the other channel and mixes with the ATC (or Cys) as they travel to the analysis sites. As the solution reaches the analysis sites, a yellow color change occurs indicating the reaction of thiol with DTNB to produce the yellow anion TNB2-. The sites are dried, scanned, yielding a linear range of inverse yellow mean intensity versus substrate concentration. An IC50 value (1.741 nM) with a known inhibitor, neostigmine bromide (NB), is obtained on the device. The multiplex design enables triplicate data collection. µTADs have great potential to be employed in a myriad of tests including point-of-care (POC) diagnostic devices for resource-challenged settings.

Keywords: paper microfluidics, acetylcholinesterase, assay

This document is currently not available here.

Share

COinS
 
Nov 12th, 10:00 AM Nov 12th, 10:15 AM

Thread-Based Microfluidic Chips as a Platform to Assess Acetylcholinesterase Activity

Watkins 2141

The quantitative detection of acetylcholinesterase (AChE) by colorimetric analysis utilizing a microfluidic thread-based analytical device (µTAD) is described. It can be difficult to study neurotransmitters (e.g. acetylcholine [ACh]) directly, and therefore the activity of AChE can be studied. Levels of ACh are relevant to Alzheimer’s disease as levels can drop by as much as 90% in patients. One way to compensate for lower levels of ACh is to inhibit the AChE enzyme from breaking down ACh. Hence, the development of facile methods to detect AChE and screen potential inhibitors will aid in the diagnosis (and eventual cure) for Alzheimer’s disease. In the present work, two µTAD platforms were fabricated. Both had a nylon thread trifurcated into three channels terminating at open sites. One platform had one end of the thread split into two (third end cut off). The nylon on the second platform was not trifurcated at the other end, creating a single inlet. 5,5’-Dithiobis-(2-nitrobenzoic acid) (DTNB) was spotted on the analysis sites. Acetylthiocholine iodide (ATC) (or cysteine, Cys), is transported through one channel of the nylon thread by capillary action due to the hydrophilic nature of nylon. AChE is transported through the other channel and mixes with the ATC (or Cys) as they travel to the analysis sites. As the solution reaches the analysis sites, a yellow color change occurs indicating the reaction of thiol with DTNB to produce the yellow anion TNB2-. The sites are dried, scanned, yielding a linear range of inverse yellow mean intensity versus substrate concentration. An IC50 value (1.741 nM) with a known inhibitor, neostigmine bromide (NB), is obtained on the device. The multiplex design enables triplicate data collection. µTADs have great potential to be employed in a myriad of tests including point-of-care (POC) diagnostic devices for resource-challenged settings.

Keywords: paper microfluidics, acetylcholinesterase, assay