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细胞功能测定

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Immobilization of Whole Cells by Chemical Vapor Deposition of Silica

Effective entrapment of whole bacterial cells onto solid-phase materials can significantly improve bioprocessing and other biotechnology applications. Cell immobilization allows integration of biocatalysts in a manner that maintains long-term cell viability and typ ...

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New Opportunities for Immobilization of Enzymes

In this chapter, as a general introduction, we summarize our personal point of view on immobilization technique in order to prepare optimal and cost-effective biocatalysts. Special attention is paid to the improvement of enzyme properties via immobilization techniques. From the sta ...

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Biomedical Applications of Immobilized Enzymes: An Update

Immobilized enzymes have been widely studied during the last few decades. Biocatalyst systems may work as biosensors or may be used for the treatment of different diseases. This chapter presents different attempts to immobilize enzymes in the biomedical field, particularly the use of pr ...

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High Activity Preparations of Lipases and Proteases for Catalysis in Low Water Containing Organic Solvents and Ionic Liquids

Simple precipitation of enzymes has shown impressive catalytic efficiencies in organic solvents. In asmuch as these can be recovered after the reaction, these can be viewed as immobilized preparations just like more extensively used cross-linked enzyme aggregates (CLEAs). This ch ...

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Improving Lipase Activity by Immobilization and Post-immobilization Strategies

One important parameter for the application of lipase catalysts in chemical industries is the specific activity displayed towards natural or unnatural substrates. Different strategies to enhance the lipase activity have been described. The immobilization of lipases on hydrop ...

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Practical Protocols for Lipase Immobilization via SolGel Techniques

Lipases can be efficiently entrapped in the pores of hydrophobic silicates by a simple and cheap sol–gel process in which a mixture of a hydrophobic alkylsilane RSi(OCH3)3 and Si(OCH3)4 is hydrolyzed under basic conditions in the presence of the enzyme. Additives such as isopropanol, polyvi ...

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Enzyme Immobilization by Entrapment Within a Gel Network

This chapter provides a detailed description of the three immobilization methods based on the biomolecules entrapment into polymer matrices. The poly (vinyl alcohol) bearing styrylpyridinium groups (PVA-SbQ), a soluble pre-polymer bearing photo-cross-linkable groups, has ...

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Modification of Carbon Nanotube Electrodes with 1-Pyrenebutanoic Acid, Succinimidyl Ester for Enhanced Bioelectrocatalysis

Conductive materials functionalized with redox enzymes provide bioelectronic architectures with application to biological fuel cells and biosensors. Effective electron transfer between the enzyme (biocatalyst) and the conductive materials is imperative for functi ...

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Immobilization of Enzymes on Ethynyl-Modified Electrodes via Click Chemistry

This paper describes a novel, simple, and versatile protocol for covalent immobilization of enzyme on electrode. The immobilization method is based on the combination of diazonium salt electrografting and click chemistry. The ethynyl-terminated monolayers are obtained by diaz ...

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Design and Characterization of Functional Nanoparticles for Enhanced Bio-performance

Recent years have witnessed the rapid development of inorganic nanomaterials for medical applications. At present, nanomedicines—nanoparticles (NPs) destined for therapy or diagnosis purposes—can be found in a number of medical applications including therapeutics (eit ...

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Tips for the Functionalization of Nanoparticles with Antibodies

Multiple antibody immobilization methodologies have been developed for several applications including affinity chromatography, immunosensing, and drug delivery. Most of them have been carried out without considering the orientation of the antigen binding site of the anti ...

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Immobilization of Enzymes on Magnetic Beads Through Affinity Interactions

The development of enzyme immobilization techniques that will not affect catalytic activity and conformation is an important research task. Affinity tags that are present or added at a specific position far from the active site in the structure of the native enzyme could be used to create stro ...

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PARP1 Genomics: Chromatin Immunoprecipitation Approach Using Anti-PARP1 Antibody (ChIP and ChIP-seq)

Poly(ADP-ribose) polymerase1 (PARP1) is a global regulator of different cellular mechanisms, ranging from DNA damage repair to control of gene expression. Since PARP1 protein and pADPr have been shown to persist in chromatin through cell cycle, they may both act as epigenetic markers. Howe ...

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Methods for Purification of Proteins Associated with Cellular Poly(ADP-Ribose) and PARP-Specific Poly(ADP-Ribose)

Poly(ADP-ribose) (pADPr) is a posttranslational modification that regulates protein function through two major mechanisms: covalent modification of acceptor proteins and noncovalent binding of proteins to pADPr. pADPr is synthesized by a family of enzymes called poly(ADP-ri ...

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Purification of Human PARP-1 and PARP-1 Domains from Escherichia coli for Structural and Biochemical Analysis

A general method to express and purify full-length human poly(ADP-ribose) polymerase-1 (PARP-1), individual PARP-1 domains, and groups of PARP-1 domains from Escherichia coli cells is described. The procedure allows for robust production of highly pure PARP-1 that is free of DNA contamin ...

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The Use of PARP Inhibitors in Cancer Therapy: Use as Adjuvant with Chemotherapy or Radiotherapy; Use as a Single Agent in Susceptible Patients; Techni

This chapter describes some of the techniques in use in our laboratories for the investigation of PARP inhibitors in clinical medicine. More specifically, we are involved in investigating the utility of PARP inhibitors in the treatment of hematopoietic malignancies. We are also active ...

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Purification of Recombinant Poly(ADP-Ribose) Polymerases

The purification of Poly(ADP-ribose) polymerases from overexpressing cells (Sf9 insect cells, Escherichia coli) has been updated to a fast and reproducible three chromatographic steps protocol. After cell lysis, proteins from the crude extract are separated on a Heparine Sepharo ...

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Detecting and Quantifying pADPr In Vivo

Poly(ADP-ribose) polymerases (PARP) participate in diverse biological processes contributing to cellular homeostasis or exacerbating injury. PARP catalyzes the addition of ADP-ribose molecules (pADPr) to the target proteins, a process termed poly-ADP-ribosylation. Ov ...

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Affinity-Based Assays for the Identification and Quantitative Evaluation of Noncovalent Poly(ADP-Ribose)-Binding Proteins

Poly(ADP-ribose) polymerases have been linked to several cellular functions, most of which being mediated through the dynamics of poly(ADP-ribose) (pADPr). In several pathways, pADPr is the effector molecule that regulates cellular signaling and dictates biological outcomes. p ...

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Noncovalent pADPr Interaction with Proteins and Competition with RNA for Binding to Proteins

PARP1 can modify a variety of proteins through conserved domains in noncovalent manner. Since poly(ADP-ribose) is highly negatively charged and has a strong binding affinity for its target proteins, noncovalent binding by poly(ADP-ribose) modulates the protein activity during dev ...

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