Aptamer entrapment in microfluidic channel using one-step sol-gel process, in view of the integration of a new selective extraction phase for lab-on-a-chip

被引:6
|
作者
Perreard, Camille [1 ,2 ,3 ,4 ]
d'Orlye, Fanny [1 ,2 ,3 ,4 ]
Griveau, Sophie [1 ,2 ,3 ,4 ]
Liu, Baohong [5 ]
Bedioui, Fethi [1 ,2 ,3 ,4 ]
Varenne, Anne [1 ,2 ,3 ,4 ]
机构
[1] PSL Res Univ, Chim ParisTech, Unite Technol Chim & Biol Sante, Paris, France
[2] INSERM, Unite Technol Chim & Biol Sante U 1022, Paris, France
[3] CNRS, Unite Technol Chim & Biol Sante, UMR 8258, Paris, France
[4] Univ Paris 05, Sorbonne Paris Cite, Unite Technol Chim & Biol Sante, Paris, France
[5] Fudan Univ, Dept Chem, Shanghai, Peoples R China
关键词
Affinity; Aptamer; Electrophoretic microchannel; Fluorescence microscopy; Retention; Sol-gel; DNA APTAMERS; SILICA; ANTIBODIES; POLY(DIMETHYLSILOXANE); ENCAPSULATION; ENZYMES; GLASS; ACID;
D O I
10.1002/elps.201600575
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
There is a great demand for integrating sample treatment into TASs. In this context, we developed a new sol-gel phase for extraction of trace compounds in complex matrices. For this purpose, the incorporation of aptamers in silica-based gel within PDMS/glass microfluidic channels was performed for the first time by a one-step sol-gel process. The effective gel attachment onto microchannel walls and aptamer incorporation in the polymerized gel were evaluated using fluorescence microscopy. A good gel stability and aptamer incorporation inside the microchannel was demonstrated upon rinsing and over storage time. The ability of gel-encapsulated aptamers to interact with its specific target (either sulforhodamine B as model fluorescent target, or diclofenac, a pain killer drug) was assessed too. The binding capacity of entrapped aptamers was quantified (in the micromolar range) and the selectivity of the interaction was evidenced. Preservation of aptamers binding affinity to target molecules was therefore demonstrated. Dissociation constant of the aptamer-target complex and interaction selectivity were evaluated similar to those in bulk solution. This opens the way to new selective on-chip SPE techniques for sample pretreatment.
引用
收藏
页码:2456 / 2461
页数:6
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