Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1291, Issue -, Pages 92-96Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2013.03.063
Keywords
Fluorescent; Microfluidic; Aptamer; Monolith
Funding
- Shandong Science and Technology Development Planning [2012GHY11510]
- [NIH-EB006124]
- [985-CXJJ201112]
- [985-CXJJ201104]
- [HIT(WH)Y201104]
- [HIT(WH)Y201103]
- [2011DXGJ15]
Ask authors/readers for more resources
A microfluidic chip with integrated 2 mm long monoliths incorporated with poly(ethylene glycol) (PEG) groups was developed for thrombin-aptamer interaction study. The non-G quartet forming oligonucleotide coated monoliths was compared to a 15 mer thrombin-binding aptamer, in which affinity binding and elution processes were real-time monitored fluorescently. The results showed that the fluorescence intensity of aptamer stationary phase is approximately 10 times higher than that of the control column, which is probably due to the successful suppression of nonspecific adsorption between thrombin and aptamers/monoliths by using PEG-monolith. The experiment was repeated using human serum albumin (HSA) and green fluorescence protein (GFP) as interferences, it was double confirmed that thrombin was selectively retained by PEG-monolith. An elution efficiency of 75% was achieved with an elute of 200 mM acetic acid and 2 M NaCl, and the eluted thrombin was successfully separated in an ionic buffer system of 20 mM NaHCO3 (pH 9.5) with 3% PEG. The hydrophilic and antifouling properties of PEG-monolith greatly decrease nonspecific adsorption and enhance detection sensitivity, which provided an alternative method to perform on-chip fluorescent measurement of bioaffinity binding. (C) 2013 Elsevier B.V. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available