Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.2.1.36 (hyaluronidase)
 4,606 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A reversed-phase ion-pair HPLC method for separating hyaluronic acid oligomers, using a polymeric C18 column at alkaline pH, is described. As the concentration of the ion-pairing agent tetrabutylammonium hydroxide increased, over the range of 0.01 to 0.06M, the capacity factors (k') of tetra- to dodecasaccharide decreased. The change in k', for each increment in pairing agent, increased with oligomer molecular weight. When changing mobile phase pH from 7 to 8, k' dramatically decreased and remained unchanged from pH 8 to 11. The isocratic separation was optimized to resolve tetrato dodecasaccharide at pH 9.0 in under 19 min. The postcolumn derivatizing agent 2-cyanoacetamide reacted with the reducing N-acetylglucosamine end groups of hyaluronic acid oligomers to yield reaction products that were monitored at 27 nm. In a series of control experiments using decasaccharide and N-acetylglucosamine, it was found that maximum product formation took place at pH 9 and was greatly influenced by borate buffer concentration. The optimum concentration for 2-cyanoacetamide was 0.33% and a temperature of 100 degrees C gave the best signal to noise ratio for the postcolumn reaction. The method is linear and reproducible, and has a lower limit of detection for tetrasaccharide of 20 ng (25 pmol). This system is suitable for studying the degradation kinetics of purified hyaluronic acid oligomers by bovine testicular hyaluronidase. Extension of the method to fluorescent and electrochemical detection and its applicability to other glycosaminoglycans is discussed.
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PMID:A reversed-phase ion-pair high-performance liquid chromatography method for bovine testicular hyaluronidase digests using postcolumn derivatization with 2-cyanoacetamide and ultraviolet detection. 188 31

The location of lectin binding sites and of anionic components was studied in the embryonic rat cerebral cortex after the formation of the cortical plate at embryonic day 18. The cortical layers advanced in differentiation, i.e. the sub-plate region and the marginal zone, showed a predominant staining with peroxidase conjugates of wheat germ agglutinin (WGA), peanut agglutinin (PNA), and after immunocytochemical detection of PNA binding sites. This pattern was obtained also with the colloidal iron hydroxide staining method. In contrast to this, the binding of concanavalin A and of succinylated WGA did not reveal a prevalent staining of the sub-plate region and the marginal zone. The further histochemical analysis of the substances responsible for the selective staining of these layers was performed by lipid extractions and by enzymatic treatment of the tissue sections with trypsin, hyaluronidase or neuraminidase prior to the binding of lectins or colloidal iron. The results obtained indicated high concentrations of sialylated galactosylglycoproteins in coexistence with glycosaminoglycans. Electron microscopy was performed with peroxidase conjugates of WGA and PNA. Binding sites of both of the lectins in the sub-plate region and in the marginal zone were located mainly at cell surfaces of the different cellular structures. The most intensive binding of WGA and PNA was detected at the surface membranes and at intracellular material of amoeboid microglial cells and astrocyte-like cell processes. It can be concluded that in distinct brain areas during early differentiation specific glycoproteins in coexistence with glycosaminoglycans are situated at, or associated with cell surfaces in high concentrations. The identical histochemical features previously described in mesenchymal tissues suggest that these glycoconjugates might be related to common morphogenetic processes in which non-neuronal cells of brain and body are specifically involved.
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PMID:Lectin binding sites and anionic components related to differentiation in the prenatal rat cerebral cortex. 384 46

Purification of proteins based on immunoaffinity has been performed using a solid support coated with antibody against the target proteins. The method requires immobilizing the antibody onto the solid support using protein A or G, and has a risk of adsorptive loss of target proteins onto the solid support. Centrifugal precipitation chromatography has been successfully used to purify enzymes, such as ketosteroid isomerase and hyaluronidase without the use of solid support. The purpose of this study is to demonstrate that immunoaffinity centrifugal precipitation chromatography is capable of isolating an antigen by exploiting antigen-antibody binding. The separation was initiated by filling both channels with 40% saturated ammonium sulfate (AS) of pH 4-4.5 followed by loading 20 microl of human plasma (National Institutes of Health blood bank) mixed with 2 mg of rabbit anti-HSA (human serum protein) antibody (Sigma). Then, the sample channel was eluted with water at 0.03 ml/min and AS channel with 40% AS solution of pH 4-4.5 at 1 ml/min until all non-binding components were eluted. Then, the releasing reagent (50% AS solution containing 0.5 M glycine and 10% ammonium hydroxide at pH 10) was introduced through the AS channel to release the target protein (HSA). The retained antibody was recovered by eluting the sample channel with water at 1 ml/min. A hollow fiber membrane device at the outlet (MicroKros, Spectrum, New Brunswick, NJ, USA) was provided on-line dialysis of the eluent before fractions were collected, so that the fractions could be analyzed by SDS-PAGE (sodium dodecyl sulfate - polyacrylamide gel electrophoresis) without further dialysis. The current method does not require immobilizing the antibody onto a matrix, which is used by the conventional immunoaffinity chromatography. This method ensures full recovery of the antigen and antibody, and it may be applied to purification of other proteins.
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PMID:Immunoaffinity centrifugal precipitation chromatography. 1741 78

Copper nanoclusters (CuNCs) have attracted considerable research interest due to their good physicochemical properties, ease of preparation, and low price. However, the low quantum yield and poor stability in aqueous solutions have greatly limited their applications. In order to improve the fluorescence properties and stability of CuNCs, in this paper, the surface confinement effect of CuNCs based on 2D layered double hydroxide (LDH) was proposed to prepare the fluorescent composites of glutathione protected CuNCs and LDH (GS-CuNCs/LDH) with excellent quantum yield and long fluorescence lifetime. Moreover, a novel, simple, and ultrasensitive fluorescence assay for the detection of hyaluronidase was proposed based on the surface confinement effect. The limit of detection for hyaluronidase was as low as 0.014 U mL-1. For the first time, this work developed a bio-enzyme sensing platform based on the surface confinement effect, which can serve as a promising candidate in biosensing.
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PMID:Investigation of the surface confinement effect of copper nanoclusters: construction of an ultrasensitive fluorescence turn-on bio-enzyme sensing platform. 3170 81

Development of unique theranostic nanoplatforms for tumor imaging and therapy remains an active topic in current nanomedicine. Here, we designed a novel targeted theranostic nanoplatform for enhanced T1 -weighted magnetic resonance (MR) imaging-guided chemotherapy by constructing layered double hydroxide (LDH)-stabilized ultrasmall iron oxide (Fe3O4) nanoparticles with hyaluronic acid (HA) modified as targeting agents, and anticancer drug doxorubicin (DOX) loaded with a high loading efficiency. Methods: The structure and release property of LDH-Fe3O4-HA/DOX nanoplatforms were characterized systematically. B16 melanoma cells with CD44 receptors overexpressed were used as model cells to determine the biocompatibility, targeting capability, and therapeutic efficiency of nanoplatforms. For in vivo experiment, hyaluronidase (HAase) pretreatment was combined with nanoplatform administration to investigate the MR imaging and chemotherapeutic effect. Results: The LDH-Fe3O4-HA nanohybrids possess good colloidal stability and cytocompatibility, display an r1 relaxivity 10-fold higher than the pristine ultrasmall Fe3O4 (4.38 mM-1 s-1 vs 0.42 mM-1 s-1), and could release drug in a pH-responsive manner. In vitro experiments demonstrate that LDH-Fe3O4-HA/DOX nanohybrids are able to specifically target B16 cells overexpressing CD44 receptors and effectively release DOX to nucleus. In vivo results show that with the pretreatment of tumor tissue by HAase to degrade the overexpressed HA in extra-cellular matrix, the designed nanoplatforms have a better tumor penetration for significantly enhanced MR imaging of tumors and tumor chemotherapy with low side effects. Conclusion: The designed LDH-Fe3O4-HA/DOX nanohybrids may be developed as a novel targeted theranostic nanoplatform for enhanced T1 -weighted MR imaging-guided chemotherapy of CD44 receptor-overexpressing tumors.
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PMID:LDH-stabilized ultrasmall iron oxide nanoparticles as a platform for hyaluronidase-promoted MR imaging and chemotherapy of tumors. 3219 35