Development of MIP-based assays for drugs of abuse
Lead Research Organisation:
University of Leicester
Department Name: Chemistry
Abstract
In this project we propose developing a novel assay for the measurement of the drugs of abuse and designer drugs which will be completely abiotic. This assay will not rely on natural antibodies and enzymes which would be replaced by magnetic and fluorescent molecularly imprinted polymer nanoparticles (MIP NPs). The proposed assay will not require refrigeration or additional procedures such as washing, elution or adding reagents, it will have long shelf and operational life, multiplexing capabilities and thus be suitable for use in the field by police force. Currently there is a gap in the market for this type of devices, especially in relations to detecting benzodiazepines. The antibodies-based assays for these compounds have shown unacceptable level of false positives. In addition development of antibodies-based sensors for designer drugs is impractical due to long period required for the production and validation of relevant antibodies. Our assay, if successful, could be used as a blueprint for the development of assays and sensors for other important target compounds and industry (MIP Diagnostics and assay manufacturers) will benefit strongly from its generic platform nature. The proposed assay will be based on a new format of the microtiter plate equipped with magnetic inserts which was developed by Leicester Biotechnology Group (GB1114231.2, and US 20150119274 A10). This technology has already been shown to work at lab scale, therefore, we believe that it is a good time to translate this technology for industrial use and apply it for detection of the compound of practical importance. The target compound for the test-system will be selected from the list of priority targets provided by Scottish Police and Crime Unit (Diazepam, Etizolam, Diclazepam, Clonazolam, Phenazepam, U-47700, 5-Fluoro-AKB48, FUB-AMB, AB-FUMINACA, JWH-018, 5-Fluoro-ADB, 5-F-MDMB-PIINACA, MMB2201, Fentanyl). The Scottish police authorities have kindly agreed to test and validate developed assay.
The project involves use of molecular modelling for selecting functional monomers which possess high affinity towards the template. Molecular modelling of MIPs is pioneered by Leicester Biotechnology Group (LBG) and relies on Sybyl 7.3 software and Leapfrog algorithm (Tripos, USA). The results of modelling will be used in polymer preparation. The training in the use of molecular modelling will be provided to student by Dr. Kal Karim.
Synthesis and characterisation of fluorescent MIP nanoparticles will be performed as described in (Canfarotta F., Poma A., Guerreiro A., Piletsky A. (2016). Solid-phase synthesis of molecularly imprinted nanoparticles (MIP NPs) for diagnostic applications. Nature Protocols, 11, 443-455).
Fluorescent MIP nanoparticles will be used as a replacement for antibodies in novel magnetic assay format developed by Leicester group (US 20150119274 A10).
The developed assay will be validated against analytical gold standards - HPLC MS. Training in using equipment will be provided by LBG members.
The project involves use of molecular modelling for selecting functional monomers which possess high affinity towards the template. Molecular modelling of MIPs is pioneered by Leicester Biotechnology Group (LBG) and relies on Sybyl 7.3 software and Leapfrog algorithm (Tripos, USA). The results of modelling will be used in polymer preparation. The training in the use of molecular modelling will be provided to student by Dr. Kal Karim.
Synthesis and characterisation of fluorescent MIP nanoparticles will be performed as described in (Canfarotta F., Poma A., Guerreiro A., Piletsky A. (2016). Solid-phase synthesis of molecularly imprinted nanoparticles (MIP NPs) for diagnostic applications. Nature Protocols, 11, 443-455).
Fluorescent MIP nanoparticles will be used as a replacement for antibodies in novel magnetic assay format developed by Leicester group (US 20150119274 A10).
The developed assay will be validated against analytical gold standards - HPLC MS. Training in using equipment will be provided by LBG members.
People |
ORCID iD |
| Sergey Piletsky (Primary Supervisor) | |
| Fabiana Grillo (Student) |
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| EP/R51326X/1 | 01/10/2018 | 30/09/2023 | |||
| 2269601 | Studentship | EP/R51326X/1 | 07/01/2019 | 30/06/2022 | Fabiana Grillo |
| Description | This award gave me the opportunity to work on a field that is in exponential growth. Drugs of abuse, and its related usage and deaths, is one of the most concerning issues worldwide. Molecularly imprinting technology was successfully applied in the development of a new sensor technology for the detection of drugs of abuse. Ongoing research of this award has the potential of becoming high impact outcome. |
| Exploitation Route | The outcomes of this funding might be used to apply for further funding and continue the current research. |
| Sectors | Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
| URL | https://doi.org/10.1016/j.bios.2021.113002 |
| Description | The current research is considerably close to industrial research it (partially already had), and will have an impact on IP generation and therefore an economic impact. |
| Sector | Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
| Impact Types | Cultural,Economic |
| Description | RSC Travel Grant (T20-4633) |
| Amount | £800 (GBP) |
| Funding ID | RSC Travel Grant (T20-4633) |
| Organisation | Royal Society of Chemistry |
| Sector | Charity/Non Profit |
| Country | United Kingdom |
| Start | 02/2020 |
| Title | Molecularly imprinted nanoparticles assay |
| Description | Molecularly imprinted polymers were synthetized for the detection of fentanyl and the polymers were applied in a novel assay platform. |
| Type Of Material | Technology assay or reagent |
| Year Produced | 2019 |
| Provided To Others? | No |
| Impact | This technology was applied for the first time for the detection of drugs of abuse. the assay developed has the potential of becoming a cheap and rapid alternative to ELISA, being one of the most common technologies currently in use for the detection of drugs. |
| Description | Carboxyl-fentanyl integrated to drugs of abuse |
| Organisation | University of Nottingham |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Some of the material synthetized by me as part of my PhD project was sent to the University of Nottingham Optics and Photonics, where it has been integrated to LPG sensor. |
| Collaborator Contribution | The production of the LPG sensor and testing of the integrated technology. |
| Impact | It is an interdisciplinary collaboration, from our front we provide polymeric chemistry and biotechnological knowledge, integrating them with their engineering, optical and photonics knowledge. The collaboration was beneficial for both partners and lead to a publication (https://doi.org/10.1016/j.bios.2021.113002) |
| Start Year | 2019 |
| Description | Integration of MIPs into nanodevice for the binding with antibodies |
| Organisation | Magna Græcia University |
| Country | Italy |
| Sector | Academic/University |
| PI Contribution | The polymers syntetised by myself were used to be integrated in a novel nanotechnological platform for the detection of multiple markers. |
| Collaborator Contribution | Providing the nanotechnological platform. |
| Impact | Chapter of my PhD thesis, one publication so far, potential patentable material. |
| Start Year | 2020 |
| Description | School presentation about my research activity |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Schools |
| Results and Impact | 30 students attended the online seminar about drugs usage and how science is helping in the fight against it. Many questions were raised from it. The event had a reasonable success not only from a scientific point of view, but also as a "inspirational talk" the students were from a different country and they asked many questions on what impact it had on my personal experience being exposed to so many different cultures in an university environment. |
| Year(s) Of Engagement Activity | 2021 |