Direct Replacement of Antibodies with Molecularly Imprinted Polymer Nanoparticles in ELISA—Development of a Novel Assay for Vancomycin
- 15 August 2013
- journal article
- research article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 85 (17), 8462-8468
- https://doi.org/10.1021/ac402102j
Abstract
A simple and straightforward technique for coating microplate wells with molecularly imprinted polymer nanoparticles (nanoMIPs) to develop assays similar to the enzyme-linked immunosorbent assay (ELISA) is presented here for the first time. NanoMIPs were synthesized by a solid-phase approach with an immobilized vancomycin (template) and characterized using Biacore 3000, dynamic light scattering, and electron microscopy. Immobilization, blocking, and washing conditions were optimized in microplate format. The detection of vancomycin was achieved in competitive binding experiments with a horseradish peroxidase–vancomycin conjugate. The assay was capable of measuring vancomycin in buffer and in blood plasma within the range of 0.001–70 nM with a detection limit of 0.0025 nM (2.5 pM). The sensitivity of the assay was 3 orders of magnitude better than a previously described ELISA based on antibodies. In these experiments, nanoMIPs have shown high affinity and minimal interference from blood plasma components. Immobilized nanoMIPs were stored for 1 month at room temperature without any detrimental effects to their binding properties. The high affinity of nanoMIPs and the lack of a requirement for cold chain logistics make them an attractive alternative to traditional antibodies used in ELISA.Keywords
This publication has 24 references indexed in Scilit:
- Rapid Determination of Metolcarb Residues in Foods Using a Biomimetic Enzyme-Linked Immunosorbent Assay Employing a Novel Molecularly Imprinted Polymer Film as Artificial AntibodyJournal of AOAC International, 2013
- DEVELOPMENT OF A BIOMIMETIC ENZYME-LINKED IMMUNOSORBENT ASSAY METHOD BASED ON A HYDROPHILIC MOLECULARLY IMPRINTED POLYMER FILM FOR DETERMINATION OF OLAQUINDOX IN CHICK FEED SAMPLESJournal of Immunoassay and Immunochemistry, 2013
- Development of a Biomimetic Enzyme-Linked Immunosorbent Assay Method for the Determination of Estrone in Environmental Water using Novel Molecularly Imprinted Films of Controlled Thickness as Artificial AntibodiesJournal of Agricultural and Food Chemistry, 2009
- Chemical Grafting of Molecularly Imprinted Homopolymers to the Surface of Microplates. Application of Artificial Adrenergic Receptor in Enzyme-Linked Assay for β-Agonists DeterminationAnalytical Chemistry, 2000
- ELISA-based assay for scatchard analysis of ligand-receptor interactionsMolecular Biotechnology, 1998
- Use of a series of homologous in vitro and in vivo assays to evaluate the endocrine modulating actions of β-sitosterol in rainbow troutAquatic Toxicology, 1998
- Imprinted Membranes for Sensor Technology: Opposite Behavior of Covalently and Noncovalently Imprinted MembranesMacromolecules, 1998
- Structure-binding relation of philanthotoxins from nicotinic acetylcholine receptor binding assayBioorganic & Medicinal Chemistry, 1997
- Mimics of the binding sites of opioid receptors obtained by molecular imprinting of enkephalin and morphine.Proceedings of the National Academy of Sciences of the United States of America, 1995
- Drug assay using antibody mimics made by molecular imprintingNature, 1993