Immunoglobulin G recognition with F_ab fragments imprinted monolithic cryogels: Evaluation of the effects of metal-ion assisted-coordination of template molecule

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• 作者：Sevgi Asliyuce^a ; Lokman Uzun^a ; ^{lokman@hacettepe.edu.tr} ; Ridvan Say^b ; Adil Denizli^a
• 关键词：Molecular imprinting ; Fab fragment ; IgG recognition ; Cu(II)-assisted coordination ; Non-covalent imprinting approaches
• 刊名：Reactive and Functional Polymers
• 出版年：2013
• 出版时间：June, 2013
• 年：2013
• 卷：73
• 期：6
• 页码：813-820
• 全文大小：1670 K

文摘

In this study, we applied the epitope imprinting approach to prepare molecularly imprinted monolithic cryogels for immunoglobulin G (IgG) recognition. In this respect, we imprinted F_ab fragments of IgG molecules instead of intact protein molecules via two different non-covalent interactions. In the first approach, we directly coordinated F_ab fragments with N-methacryloyl-L-histidine (MAH), polymerizable derivative of L-histidine, but for the second, we used cupric ions [Cu(II)] as mediator between MAH and F_ab fragments. The monolithic cryogels were characterized by Fourier transform infrared (FTIR) spectroscopy, swelling test, and scanning electron microscopy. Then, the monolithic cryogels were used for F_ab fragment adsorption from aqueous solution while evaluating the factors such as pH and F_ab fragment concentration affecting on adsorption process in continuous set-up. After that, monolithic cryogels were used for IgG adsorption by varying pH, IgG concentration, flowrate, and temperature in appropriate ranges. Maximum IgG adsorption capacities were determined as 32.4 mg/g and 49.0 mg/g for directly coordinated cryogel (MIP_Direct) and Cu(II) assisted cryogel (MIP_{Cu(II) assisted}), respectively. Non-imprinted monolithic cryogels were also prepared for control purposes. In addition to F_ab fragments and IgG molecules, albumin and F_c fragment of IgG molecules were used as competitor biomolecules in order to investigate the selectivity gained by imprinting process. Relative selectivity constants were calculated as 1.47, 2.64 and 3.89 for MIP_Direct and 2.90, 8.98, and 11.51 for MIP_{Cu(II) assisted} for F_ab/IgG, F_ab/F_c, and F_ab/albumin as biomolecule pairs, respectively. The desorption efficiency and reusability of MIP_{Cu(II) assisted} cryogel were better than that of MIP_Direct. The results reported here showed that the metal ion assistance improved the selectivity features of the imprinted cryogels and allowed to study under milder conditions with enhanced adsorptive properties.