| 1. |
- Contardi, Cecilia, et al.
(författare)
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Affinity Capillary Electrophoresis as a Tool To Characterize Molecularly Imprinted Nanogels in Solution
- 2024
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Ingår i: Analytical Chemistry. - : American Chemical Society (ACS). - 0003-2700 .- 1520-6882. ; 96:7, s. 3017-3024
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Tidskriftsartikel (refereegranskat)abstract
- In this work, an innovative and accurate affinity capillary electrophoresis (ACE) method was set up to monitor the complexation of aqueous MIP nanogels (NGs) with model cancer-related antigens. Using α2,6′- and α2,3′-sialyllactose as oversimplified cancer biomarker-mimicking templates, NGs were synthesized and characterized in terms of size, polydispersity, and overall charge. A stability study was also carried out in order to select the best storage conditions and to ensure product quality. After optimization of capillary electrophoresis conditions, injection of MIP NGs resulted in a single, sharp, and efficient peak. The mobility shift approach was applied to quantitatively estimate binding affinity, in this case resulting in an association constant of K ≈ 106 M–1. The optimized polymers further displayed a pronounced discrimination between the two sialylated sugars. The newly developed ACE protocol has the potential to become a very effective method for nonconstrained affinity screening of NG in solution, especially during the NG development phase and/or for a final accurate quantitation of the observed binding.
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| 2. |
- Contardi, Cecilia, et al.
(författare)
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Rational Design of Highly Selective Sialyllactose-Imprinted Nanogels
- 2024
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Ingår i: Chemistry - A European Journal. - : John Wiley & Sons. - 0947-6539 .- 1521-3765. ; 30:45
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Tidskriftsartikel (refereegranskat)abstract
- We describe a facile method to prepare water-compatible molecularly imprinted polymer nanogels (MIP NGs) as synthetic antibodies against target glycans. Three different phenylboronic acid (PBA) derivatives were explored as monomers for the synthesis of MIP NGs targeting either α2,6- or α2,3-sialyllactose, taken as oversimplified models of cancer-related sT and sTn antigens. Starting from commercially available 3-acrylamidophenylboronic acid, also its 2-substituted isomer and the 5-acrylamido-2-hydroxymethyl cyclic PBA monoester derivative were initially evaluated by NMR studies. Then, a small library of MIP NGs imprinted with the α2,6-linked template was synthesized and tested by mobility shift Affinity Capillary Electrophoresis (msACE), to rapidly assess an affinity ranking. Finally, the best monomer 2-acrylamido PBA was selected for the synthesis of polymers targeting both sialyllactoses. The resulting MIP NGs display an affinity constant≈106 M−1 and selectivity towards imprinted glycans. This general procedure could be applied to any non-modified carbohydrate template possessing a reducing end.
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| 3. |
- Emgenbroich, Marco, et al.
(författare)
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A Phosphotyrosine-Imprinted Polymer Receptor for the Recognition of Tyrosine Phosphorylated Peptides
- 2008
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Ingår i: Chemistry - A European Journal. - : Wiley. - 0947-6539 .- 1521-3765. ; 14:31, s. 9516-29
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Tidskriftsartikel (refereegranskat)abstract
- Hyperphosphorylation at tyrosine is commonly observed in tumor proteomes and, hence, specific phosphoproteins or phosphopeptides could serve as markers useful for cancer diagnostics and therapeutics. The analysis of such targets is, however, a challenging task, because of their commonly low abundance and the lack of robust and effective preconcentration techniques. As a robust alternative to the commonly used immunoaffinity techniques that rely on phosphotyrosine(pTyr)-specific antibodies, we have developed an epitope-imprinting strategy that leads to a synthetic pTyr-selective imprinted polymer receptor. The binding site incorporates two monourea ligands placed by preorganization around a pTyr dianion template. The tight binding site displayed good binding affinities for the pTyr template, in the range of that observed for corresponding antibodies, and a clear preference for pTyr over phosphoserine (pSer). In further analogy to the antibodies, the imprinted polymer was capable of capturing short tyrosine phosphorylated peptides in the presence of an excess of their non-phosphorylated counterparts or peptides phosphorylated at serine.
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| 4. |
- Oxelbark, Joakim, et al.
(författare)
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Chromatographic comparison of bupivacaine imprinted polymers prepared in crushed monolith, microsphere, silica-based composite and capillary monolith formats
- 2007
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Ingår i: Journal of Chromatography A. - : Elsevier BV. - 0021-9673. ; 1160:1-2, s. 215-26
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Tidskriftsartikel (refereegranskat)abstract
- A comprehensive comparison of five chromatographic stationary phases based on molecularly imprinted polymers is presented. Efficiency, imprinting factors, water compatibility and batch-to-batch reproducibility are discussed for crushed monolith, microspheres, two silica-based composites and capillary monoliths, all imprinted with the local anaesthetic bupivacaine. Synthesis protocol and chromatographic test conditions have been kept fixed within certain limits, in order to provide further insight into the strengths and weaknesses of the different formats. Excluding microparticles, all formats give satisfactory performance, especially in aqueous mobile phases. An assessment of batch-to-batch reproducibility in different mobile phases adds further value to this comparison study.
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