A novel assay format as an alternative to ELISA: MINA test for biotin

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Title: A novel assay format as an alternative to ELISA: MINA test for biotin
Author(s): Piletsky, SS
Cass, AEG
Piletska, EV
Czulak, J
Piletsky, SA
Item Type: Journal Article
Abstract: A novel abiotic assay based on biotin‐specific fluorescent molecularly imprinted polymer nanoparticles (nanoMIPs) which acted as both reporter probes and binding agents, was developed. This is a first report of an assay which, unlike ELISA, required no washing steps or addition of enzyme substrates, making it more user‐friendly. The components of the molecularly imprinted polymer nanoparticles assay (MINA) were assembled in microtiter plates fitted with magnetic inserts. The fluorescent nanoMIPs were bound to biotin‐conjugated magnetic particles, which were attracted to the inserts. The addition of free biotin caused a displacement of the fluorescent nanoMIPs into solution, generating a signal proportional to the concentration of biotin. The nanoMIPs had a dissociation constant (Kd) of 14 nM, allowing the assay to detect biotin at nano‐molar concentrations. The pre‐assembled assay only required the addition of the sample and measurement of the fluorescence, and it functioned well after six weeks of storage without refrigeration. The assay did not show the susceptibility to several compounds which are known to interfere with avidin and streptavidin‐based assays, such as mercaptoethanol and sugars. The protocols optimized in this work could be used to develop the abiotic assays for any other compound of interest.
Publication Date: 1-Dec-2018
Date of Acceptance: 12-Sep-2018
URI: http://hdl.handle.net/10044/1/63465
DOI: https://dx.doi.org/10.1002/cnma.201800393
ISSN: 2199-692X
Publisher: Wiley
Start Page: 1214
End Page: 1222
Journal / Book Title: ChemNanoMat
Volume: 4
Issue: 12
Copyright Statement: © 2018 Wiley‐VCH Verlag GmbH & Co. KGaA, Weinheim. This is the accepted version of the following article: Piletsky, S. S., Cass, A. E., Piletska, E. V., Czulak, J. and Piletsky, S. A. (2018), A Novel Assay Format as an Alternative to ELISA: MINA Test for Biotin. ChemNanoMat. . doi:10.1002/cnma.201800393, which has been published in final form at https://dx.doi.org/10.1002/cnma.201800393
Keywords: Science & Technology
Physical Sciences
Chemistry, Multidisciplinary
Nanoscience & Nanotechnology
Materials Science, Multidisciplinary
Science & Technology - Other Topics
Materials Science
molecular imprinting
molecularly imprinted polymer nanoparticles
abiotic assays
Publication Status: Published
Embargo Date: 2019-10-01
Online Publication Date: 2018-10-01
Appears in Collections:Chemistry
Biological and Biophysical Chemistry
Faculty of Natural Sciences

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