Label-free Protein Detection Based on the Heat-Transfer Method--A Case Study with the Peanut Allergen Ara h 1 and Aptamer-Based Synthetic Receptors.
Aptamers are an emerging class of molecules that, because of the development of the systematic evolution of ligands by exponential enrichment (SELEX) process, can recognize virtually every target ranging from ions, to proteins, and even whole cells. Although there are many techniques capable of detecting template molecules with aptamer-based systems with high specificity and selectivity, they lack the possibility of integrating them into a compact and portable biosensor setup. Therefore, we will present the heat-transfer method (HTM) as an interesting alternative because this offers detection in a fast and low-cost manner and has the possibility of performing experiments with a fully integrated device. This concept has been demonstrated for a variety of applications including DNA mutation analysis and screening of cancer cells. To the best our knowledge, this is the first report on HTM-based detection of proteins, in this case specifically with aptamer-type receptors. For proof-of-principle purposes, measurements will be performed with the peanut allergen Ara h 1 and results indicate detection limits in the lower nanomolar regime in buffer liquid. As a first proof-of-application, spiked Ara h 1 solutions will be studied in a food matrix of dissolved peanut butter. Reference experiments with the quartz-crystal microbalance will allow for an estimate of the areal density of aptamer molecules on the sensor-chip surface.
†Institute for Materials Research, Hasselt University, Wetenschapspark 1, 3590 Diepenbeek, Belgium.|‡School of Biological and Chemical Sciences, Queen Mary University of London, Mile End Road, London E1 4NS, United Kingdom.
Marloes
M
Peeters
§Maastricht Science Programme, Maastricht University, 6200 MD Maastricht, The Netherlands.
Bart
B
van Grinsven
§Maastricht Science Programme, Maastricht University, 6200 MD Maastricht, The Netherlands.
Thomas J
TJ
Cleij
†Institute for Materials Research, Hasselt University, Wetenschapspark 1, 3590 Diepenbeek, Belgium.
Kathia Lorena
KL
Jiménez-Monroy
∥Department of Physics and Astronomy, Soft-Matter and Biophysics Section, Catholic University Leuven, Celestijnenlaan 200 D, 3001 Leuven, Belgium.
Peter
P
Cornelis
#BIOSYST-MeBioS, KU Leuven-University of Leuven, Willem de Crooylan 42, 3000 Leuven, Belgium.
Elena
E
Pérez-Ruiz
†Institute for Materials Research, Hasselt University, Wetenschapspark 1, 3590 Diepenbeek, Belgium.|∥Department of Physics and Astronomy, Soft-Matter and Biophysics Section, Catholic University Leuven, Celestijnenlaan 200 D, 3001 Leuven, Belgium.
Gideon
G
Wackers
†Institute for Materials Research, Hasselt University, Wetenschapspark 1, 3590 Diepenbeek, Belgium.|⊥Division IMOMEC, IMEC v.z.w. , Wetenschapspark 1, 3590 Diepenbeek, Belgium.
Ronald
R
Thoelen
†Institute for Materials Research, Hasselt University, Wetenschapspark 1, 3590 Diepenbeek, Belgium.|⊥Division IMOMEC, IMEC v.z.w. , Wetenschapspark 1, 3590 Diepenbeek, Belgium.
Ward
W
De Ceuninck
#BIOSYST-MeBioS, KU Leuven-University of Leuven, Willem de Crooylan 42, 3000 Leuven, Belgium.
Jeroen
J
Lammertyn
†Institute for Materials Research, Hasselt University, Wetenschapspark 1, 3590 Diepenbeek, Belgium.|∥Department of Physics and Astronomy, Soft-Matter and Biophysics Section, Catholic University Leuven, Celestijnenlaan 200 D, 3001 Leuven, Belgium.
Patrick
P
Wagner
D052179:Antigens, Plant / Q000032:analysis* / Q000737:chemistry; D052157:Aptamers, Nucleotide / Q000737:chemistry*; D015374:Biosensing Techniques / Q000379:methods*; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry; D006358:Hot Temperature; D008565:Membrane Proteins; D010940:Plant Proteins / Q000032:analysis* / Q000737:chemistry; D040081:Protein Array Analysis / Q000295:instrumentation*; D062165:Receptors, Artificial / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013194:Staining and Labeling; D013817:Thermography / Q000379:methods*
D052179:Antigens, Plant; D052157:Aptamers, Nucleotide; C500889:Ara h 1 protein, Arachis hypogaea; D006023:Glycoproteins; D008565:Membrane Proteins; D010940:Plant Proteins; D062165:Receptors, Artificial
Ara h 1; aptamers; biomimetic sensors; heat-transfer method (HTM); label-free biosensors; proteins