Chianella Iva, Lotierzo Manuela, Piletsky Sergey A, Tothill Ibtisam E, Chen Beining, Karim Khalku, Turner Anthony P F
Institute of BioScience and Technology, Cranfield University, Silsoe, Bedfordshire, UK.
Anal Chem. 2002 Mar 15;74(6):1288-93. doi: 10.1021/ac010840b.
A computational approach for the design of a molecularly imprinted polymer (MIP) specific for Cyanobacterial toxin microcystin-LR is presented. By using molecular modeling software, a virtual library of functional monomers was designed and screened against the target toxin, employed as a template. The monomers giving the highest binding energy were selected and used in a simulated annealing (molecular dynamics) process to investigate their interaction with the template. The stoichiometric ratio observed from the simulated annealing study was used in MIP preparation for microcystin-LR. The monomers were copolymerized with a cross-linker in the presence of the template. A control (blank) polymer was prepared under the same conditions but in the absence of template. A competitive assay with microcystin-horseradish peroxidase conjugate was optimized and used to evaluate the affinity and cross-reactivity of the polymer. The performance of the artificial receptor was compared to the performance of monoclonal and polyclonal antibodies raised against the toxin. The results indicate that imprinted polymer has affinity and sensitivity comparable to those of polyclonal antibodies (the detection limit for microcystin-LR using the MIP-based assay was found to be 0.1 microg L-1), while superior chemical and thermal stabilities were obtained. Moreover, cross-reactivity to other toxin analogues was very low for the imprinted polymer, in contrast to the results achieved for antibodies. It is anticipated that the polymer designed could be used in assays, sensors, and solid-phase extraction.
本文介绍了一种用于设计针对蓝藻毒素微囊藻毒素-LR的分子印迹聚合物（MIP）的计算方法。通过使用分子建模软件，设计了一个功能单体虚拟库，并针对用作模板的目标毒素进行筛选。选择给出最高结合能的单体，并用于模拟退火（分子动力学）过程，以研究它们与模板的相互作用。从模拟退火研究中观察到的化学计量比用于微囊藻毒素-LR的MIP制备。单体在模板存在下与交联剂共聚。在相同条件下但在无模板的情况下制备对照（空白）聚合物。优化了与微囊藻毒素-辣根过氧化物酶缀合物的竞争测定，并用于评估聚合物的亲和力和交叉反应性。将人工受体的性能与针对该毒素产生的单克隆和多克隆抗体进行比较。结果表明，印迹聚合物具有与多克隆抗体相当的亲和力和灵敏度（使用基于MIP的测定法对微囊藻毒素-LR的检测限为0.1μg L-1），同时获得了优异的化学和热稳定性。此外，与抗体的结果相反，印迹聚合物对其他毒素类似物的交叉反应性非常低。预计设计的聚合物可用于分析、传感器和固相萃取。
