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Oriented immobilization of PNgase F on a porous polymer monolith for rapid N-glycan release
Szekrényes, A. ; Křenková, Jana ; Keresztessy, Z. ; Foret, František ; Guttman, A.
We demonstrate a simple and rapid method for the oriented immobilization of peptide-N4-(Nacetyl- glucosaminyl) asparagine amidase F (PNGase F) on a porous polymer monolith. The oriented immobilization is based on the affinity of glutathione-S-transferase (GST) tagged PNGase F towards glutathione modified monolith prepared in the capillary format. This approach allows the oriented and easily replaceable immobilization of PNGase F for rapid and efficient release of N-linked glycans. The reactor was tested by deglycosylation of several native glycoproteins such as ribonuclease B, fetuin or human immunoglobulin G. The proteins were effectively deglycosylated in several minutes and the released N-linked glycans were analyzed using off-line MALDI/MS or CE/LIF.
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Thin metal films in resistivity-based chemical sensing
Podešva, Pavel ; Foret, František
Measurement of basic electric properties (current, voltage, resistance) is the most frequent technique in practically all scientific disciplines. Direct electric measurements are simple, precise and provide very large dynamic range. Since the related equipment is simple and inexpensive sensors based on electric measurements are very attractive for many applications in chemistry, environmental sciences or medicine. In this work, we present resistivity sensor for direct measurement of thiols in liquid samples.
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Filtration microcartridge and capillary isoelectric focusing for the analysis low number of microorganisms in real samples
Kubesová, Anna ; Horká, Marie ; Šalplachta, Jiří ; Horký, J.
At present, detection and identification of microorganisms in real biological samples needs very sensitive methods for correct diagnosis and therapy in medical practice, biotechnology, agriculture, food safety and quality etc. Accordingly, it is essential to develop new cheap techniques for pathogen identification, particularly for pathogen in complex biological samples. Pre-concentration, separation, and sensitive detection of whole cells in one step are great potential and advantage for detection of pathogens in low concentration. Combination of filtration microcartridge and capillary isoelectric focusing was developed for pre-concentration and pre-separation of microorganisms from real suspensions and the possibility its application to real samples was verified.
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