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Characterization of paramagnetic microparticles suitable for amino acids/peptides binding and their encapsulation into liposome as new carriers

The multidisciplinary field of nanotechnologies has engulfed a myriad of new opportunities in many areas of interest like health care, material sciences or analytical chemistry (Sahoo et al. 2007). In analytical chemistry, analytical processes generally consist of few basic steps, including sample preparation, detection and data treatment, to obtain proper results. Among all these procedures, sample preparation is the most important step, especially for detecting analytes in various matrices (Ravelo-Perez et al. 2010; Ouyang et al. 2011). For target analytes preconcentration, a lot of adsorption and absorption techniques have been used such as solid-phase extraction (SPE) and/or solid-phase micro extraction (SPME). It is not surprising that nanomaterials-based separation and preconcentration play important roles in many analytical procedures to concentrate analytes or to eliminate the interfering substances (Chun et al. 2011; Pompa et al. 2006). Moreover, another advantage of their utilization is modification of these materials with various chemical groups to increase their selectivity, making them applicable for extraction of analytes in complex heterogeneous matrixes (Wu et al. 2011; Liu 2009). Magnetic nanoparticles increasingly used as the special immobilizing carrier of biomolecules are currently of great interest in current researches mainly due to their biocompatibility, easily renewability, and stability against degradation (Fang and Zhang 2009; Li et al. 2011; Son et al. 2005; Insin et al. 2008). In biological sciences paramagnetic particles have been utilized for several different purposes such as transporting, carrying or for collecting of biomolecules in solution (Pettersson et al. 2013; Drbohlavova et al. 2013). The versatility of paramagnetic particles challenges to expand possibilities of their utilization and develop new combination improving their excellent qualities (Pinzon-Daza et al. 2013).

Thus, this study focuses on the synthesis and characterization of different microparticles modified with nanomaghemite providing perfect paramagnetic properties. Our goal was to develop paramagnetic microparticles (PMPs) able to bind amino acids (proline, cysteine, serine, leucine, sarcosine, glycine, valine and arginine) or peptides (reduced and oxidized glutathione) and thus be able to serve as a preconcentration tool usable to isolate various analytes and/or to immobilize target substance and thus serve as a transporter. We carried out characterization using different methods as X-ray fluorescence (XRF) providing us information about elemental composition of our particles, scanning electron microscopy (SEM) allowing us to gain some further insight into particles morphology and scanning electrochemical microscopy (SECM) evaluating the capability of paramagnetic particle surface to change its relative current in response to binding with amino acid/peptide. Further, ion-exchange liquid chromatography was used to obtain the information about the specifity of PMPs to the presence of different amino acids/peptides. Finally we enclosed the PMPs with bound amino acids into liposome modified with gold and linker with magnetic particle, which increased paramagnetic properties of the formed transporter.

We synthesized paramagnetic microparticles exhibiting different binding specifities. Using SEM, SECM, XRF and IELC methods we performed characterization showing that our PMPs are able to establish binding with different amino acids such as valine, leucine, sarcosine, arginine or peptides in the case of GSH and GSSG and thus may serve as a tool for various applications. Furthermore our next novelty was approach using the combination of liposome modified with gold nanoparticles further modified with ODT linked to gold nanoparticles via linker formed in process of triple hybridization. Finally we enclosed paramagnetic microparticles carrying required amino acid; in our case valine and leucine, into liposome´s cavity. Complex formed in this manner may serve as a tool for target transporting of different biomolecules into desired location in various tissues. Moreover, safe application is assured by a biocompatibility of individual components constituting the complex.

Práce je spojená s projektem CEITEC CZ.1.05/1.1.00/02.0068.

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