Development of MIP-based assays for drugs of abuse

开发基于 MIP 的药物滥用检测方法

• 批准号: 2269601
• 金额: --
• 依托单位: University of Leicester
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2269601
• 关键词: Development; MIP; based; assays; drugs

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.

在这个项目中，我们建议开发一种新的方法来测量药物滥用和设计药物，这将是完全非生物的。该测定将不依赖于天然抗体和酶，其将被磁性和荧光分子印迹聚合物纳米颗粒（MIP NPs）取代。所提出的测定不需要冷藏或额外的程序，如洗涤、洗脱或添加试剂，它将具有较长的保质期和操作寿命，多路复用能力，因此适合警察部队在现场使用。目前，市场上这种类型的设备存在空白，特别是在检测苯二氮卓类药物方面。这些化合物的基于抗体的测定显示出不可接受的假阳性水平。此外，由于相关抗体的生产和验证需要很长的时间，因此开发用于设计药物的基于抗体的传感器是不切实际的。如果成功的话，我们的检测方法可以作为开发其他重要目标化合物的检测方法和传感器的蓝图，行业（MIP诊断和检测制造商）将从其通用平台性质中受益匪浅。拟定试验将基于莱斯特生物技术集团开发的配备磁性插入物的新型微量滴定板（GB 1114231.2和US 20150119274 A10）。该技术已被证明在实验室规模下有效，因此，我们认为现在是将该技术转化为工业用途并将其应用于检测具有实际重要性的化合物的好时机。测试系统的目标化合物将从苏格兰警察和犯罪单位提供的优先目标列表中选择（地西泮、依替唑仑、地克拉西泮、氯硝安定、非那西泮、U-47700、5-氟-AKB48、FUB-AMB、AB-FUMINACA、JWH-018、5-氟-ADB、5-F-MDMB-PIINACA、MMB 2201、芬太尼）。苏格兰警察当局已同意测试和验证开发的检测方法。该项目涉及使用分子建模来选择对模板具有高亲和力的功能单体。MIP的分子建模由莱斯特生物技术集团（LBG）开创，并依赖于Syphony 7.3软件和Leapfrog算法（Tripos，USA）。模拟结果将用于聚合物的制备。Kal Karim博士将为学生提供使用分子建模的培训。荧光MIP纳米颗粒的合成和表征将如（Canfarotta F.，Poma A.，Guerreiro A.，皮列茨基湾（2016年）。用于诊断应用的分子印迹纳米颗粒（MIP NPs）的固相合成。Nature Protocols，11，443-455）。荧光MIP纳米颗粒将在由莱斯特组（US 20150119274 A10）开发的新型磁性测定形式中用作抗体的替代物。开发的测定将针对分析金标准- HPLC MS进行验证。LBG成员将提供使用设备的培训。

项目成果

Enhancing Antibodies' Binding Capacity through Oriented Functionalization of Plasmonic Surfaces.

• DOI: 10.3390/nano11102620
• 发表时间: 2021-10-05
• 期刊: Nanomaterials (Basel, Switzerland)
• 作者: Coluccio ML;Grillo F;Onesto V;Garo V;Scala C;Cuzzola P;Calfa M;Candeloro P;Gentile F;Piletsky S;Malara N
• 通讯作者: Malara N

Carboxyl-fentanyl detection using optical fibre grating-based sensors functionalised with molecularly imprinted nanoparticles

• DOI: 10.1016/j.bios.2021.113002
• 发表时间: 2021-01-21
• 期刊: BIOSENSORS & BIOELECTRONICS
• 影响因子: 12.6
• 作者: Liu, LiangLiang;Grillo, Fabiana;Korposh, Serhiy
• 通讯作者: Korposh, Serhiy