Search results for: linker

Authors: Hui Wang, Harish Padmanabhan, David Thomson, Krassen Dimitrov

Abstract:

Two optimized strategies were successfully established to develop biomolecule-based magnetic nanoassemblies. Streptavidin-coated and amine-coated magnetic nanoparticles were chosen as model scaffolds onto which double-stranded DNA and human immunoglobulin G were specifically conjugated in succession, using biotin-streptavidin interaction or covalent cross-linkers. The success of this study opens the prospect of developing selective and sensitive nanoparticle-based structures for diagnostics or drug delivery.

Keywords: Antibody, DNA, linker, magnetic nanoparticles, streptavidin

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Authors: Srinivas Bathini, Duraichelvan Raju, Simona Badilescu, Muthukumaran Packirisamy

Abstract:

A bio-sensing method, based on the plasmonic property of gold nano-islands, has been developed for detection of exosomes in a clinical setting. The position of the gold plasmon band in the UV-Visible spectrum depends on the size and shape of gold nanoparticles as well as on the surrounding environment. By adsorbing various chemical entities, or binding them, the gold plasmon band will shift toward longer wavelengths and the shift is proportional to the concentration. Exosomes transport cargoes of molecules and genetic materials to proximal and distal cells. Presently, the standard method for their isolation and quantification from body fluids is by ultracentrifugation, not a practical method to be implemented in a clinical setting. Thus, a versatile and cutting-edge platform is required to selectively detect and isolate exosomes for further analysis at clinical level. The new sensing protocol, instead of antibodies, makes use of a specially synthesized polypeptide (Vn96), to capture and quantify the exosomes from different media, by binding the heat shock proteins from exosomes. The protocol has been established and optimized by using a glass substrate, in order to facilitate the next stage, namely the transfer of the protocol to a microfluidic environment. After each step of the protocol, the UV-Vis spectrum was recorded and the position of gold Localized Surface Plasmon Resonance (LSPR) band was measured. The sensing process was modelled, taking into account the characteristics of the nano-island structure, prepared by thermal convection and annealing. The optimal molar ratios of the most important chemical entities, involved in the detection of exosomes were calculated as well. Indeed, it was found that the results of the sensing process depend on the two major steps: the molar ratios of streptavidin to biotin-PEG-Vn96 and, the final step, the capture of exosomes by the biotin-PEG-Vn96 complex. The microfluidic device designed for sensing of exosomes consists of a glass substrate, sealed by a PDMS layer that contains the channel and a collecting chamber. In the device, the solutions of linker, cross-linker, etc., are pumped over the gold nano-islands and an Ocean Optics spectrometer is used to measure the position of the Au plasmon band at each step of the sensing. The experiments have shown that the shift of the Au LSPR band is proportional to the concentration of exosomes and, thereby, exosomes can be accurately quantified. An important advantage of the method is the ability to discriminate between exosomes having different origins.

Keywords: Exosomes, gold nano-islands, microfluidics, plasmonic biosensing.

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Authors: Aifa Afirah Rozlan, Mohamad Suhaimi Jaafar, Azhar Abdul Rahman

Abstract:

Raman spectroscopy are used to characterize the chemical changes in normoxic polyhydroxyethylacrylate gel dosimeter (PHEA) induced by radiation. Irradiations in the low dose region are performed and the polymerizations of PHEA gels are monitored by the observing the changes of Raman shift intensity of the carbon covalent bond of PHEA originated from both monomer and the cross-linker. The variation in peak intensities with absorbed dose was observed. As the dose increase, the peak intensities of covalent bond of carbon in the polymer gels decrease. This point out that the amount of absorbed dose affect the polymerization of polymer gels. As the absorbed dose increase, the polymerizations also increase. Results verify that PHEA gel dosimeters are sensitive even in lower dose region.

Keywords: normoxic polymer gel, ascorbic acid, Ramanspectroscopy, radiation dosimetry.

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Authors: Simona Dostalova, Pavel Kopel, Marketa Vaculovicova, Vojtech Adam, Rene Kizek

Abstract:

To ensure targeting of apoferritin nanocarrier with encapsulated doxorubicin drug, we used a peptide linker based on a protein G with N-terminus affinity towards Fc region of antibodies. To connect the peptide to the surface of apoferritin, the C-terminus of peptide was made of cysteine with affinity to gold. The surface of apoferritin with encapsulated doxorubicin (APODOX) was coated either with gold nanoparticles (APODOX-Nano) or gold(III) chloride hydrate reduced with sodium borohydride (APODOX-HAu). The reduction with sodium borohydride caused a loss of doxorubicin fluorescent properties and probably accompanied with the loss of its biological activity. Fluorescent properties of APODOX-Nano were similar to the unmodified APODOX; therefore it was more suited for the intended use. To evaluate the specificity of apoferritin modified with antibodies, ELISA-like method was used with the surface of microtitration plate wells coated by the antigen (goat anti-human IgG antibodies). To these wells, the nanocarrier was applied. APODOX without the modification showed 5× lower affinity to the antigen than APODOX-Nano modified gold and targeting antibodies (human IgG antibodies).

Keywords: Antibody targeting, apoferritin, doxorubicin, nanocarrier.

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Authors: A. Deręgowska, J. Depciuch, R. Wojnarowska, J. Polit, D. Broda, H. Nechai, M. Gonchar, and E. Sheregii

Abstract:

16-Mercaptohexadecanoic acid (MHDA) and tripeptide glutathione conjugated with gold nanoparticles (Au-NPs) are characterized by Fourier Transform InfaRared (FTIR) spectroscopy combined with Surface-enhanced Raman scattering (SERS) spectroscopy. Surface Plasmon Resonance (SPR) technique based on FTIR spectroscopy has become an important tool in biophysics, which is perspective for the study of organic compounds. FTIR-spectra of MHDA shows the line at 2500 cm-1 attributed to thiol group which is modified by presence of Au-NPs, suggesting the formation of bond between thiol group and gold. We also can observe the peaks originate from characteristic chemical group. A Raman spectrum of the same sample is also promising. Our preliminary experiments confirm that SERS-effect takes place for MHDA connected with Au-NPs and enable us to detected small number (less than 106 cm-2) of MHDA molecules. Combination of spectroscopy methods: FTIR and SERS – enable to study optical properties of Au- NPs and immobilized bio-molecules in context of a bio-nano-sensors.

Keywords: Glutathione; gold nanoparticles, Fourier transform infrared spectroscopy, MHDA, surface-enhanced Raman scattering.

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