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    26390 results match your criteria Analytical Biochemistry [Journal]

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    Plasmid DNA purification by zirconia magnetic nanocomposite.
    Anal Biochem 2017 Oct 7;539:33-38. Epub 2017 Oct 7.
    Department of Biotechnology, College of Agriculture, Isfahan University of Technology, Isfahan 84156-83111, Iran.
    A zirconia magnetic nanocomposite was prepared by a co-precipitation method in one step. The synthesized nanocomposite was characterized by scanning electron microscopy, energy-dispersive X-ray and Fourier transform infrared spectroscopy. The nanocomposite was applied for the adsorption/desorption of tobacco plant DNA (as a model). Read More

    Application of RT-PCR and MALDI-TOF MS for the detection of RNA luteovirus.
    Anal Biochem 2017 Oct 6;539:45-47. Epub 2017 Oct 6.
    National Agriculture and Food Research Organization, Kannondai 3-1-1, Tsukuba, Ibaraki 305-8517, Japan.
    There is a need for rapid and less expensive methods to identify RNA viruses, including luteoviruses, for practical use in agriculture and quarantine. The mass spectrometric cleaved amplified polymorphic sequence (MS-CAPS) method, which detects enzymatically cleaved amplicons by matrix-assisted laser desorption/ionization mass spectrometry, was herein used together with a short RT-PCR to detect luteovirus in only 90 min. In addition, the matrixes 2',4',6'-trihydroxyacetophene and 3-hydroxypicolinic acid were compared for their effectiveness in the analysis of short single-stranded biotinylated DNA obtained by a MS-CAPS reaction. Read More

    Rapid quantification of glutaminase 2 (GLS2)-related metabolites by HILIC-MS/MS.
    Anal Biochem 2017 Oct 6;539:39-44. Epub 2017 Oct 6.
    School of Pharmacy, College of Medicine, National Taiwan University, Taipei, Taiwan; The Metabolomics Core Laboratory, Center of Genomic Medicine, National Taiwan University, Taipei, Taiwan; Department of Pharmacy, National Taiwan University Hospital, Taipei, Taiwan. Electronic address:
    Glutamine, glutamate and glutathione are key modulators of excessive oxidative stress in tumor cells. In this study, we developed a rapid and accurate HILIC-MS/MS method to simultaneously determine concentrations of cellular glutamine, glutamate and glutathione. A bared silica HILIC column was employed to analyze these polar metabolites. Read More

    In situ measurements of mitochondrial matrix enzyme activities using plasma and mitochondrial membrane permeabilization agents.
    Anal Biochem 2017 Oct 4. Epub 2017 Oct 4.
    University of California, San Diego, Department of Pharmacology, 9500 Gilman Drive #0636, La Jolla, CA 92093, United States.
    Activities of enzymes localized to the mitochondrial matrix of mammalian cells are often critical regulatory steps in cellular metabolism. As such, measurement of matrix enzyme activities in response to genetic modifications or drug interventions is often desired. However, measurements in intact cells are often hampered by the presence of other isozymes in the cytoplasm as well as the inability to deliver enzyme substrates across cellular membranes. Read More

    DNA interaction with platinum-based cytostatics revealed by DNA sequencing.
    Anal Biochem 2017 Sep 29;539:22-28. Epub 2017 Sep 29.
    Department of Chemistry and Biochemistry, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno, Czech Republic; Central European Institute of Technology, Brno University of Technology, Purkynova 123, CZ-612 00 Brno, Czech Republic. Electronic address:
    The main mechanism of action of platinum-based cytostatic drugs - cisplatin, oxaliplatin and carboplatin - is the formation of DNA cross-links, which restricts the transcription due to the disability of DNA to enter the active site of the polymerase. The polymerase chain reaction (PCR) was employed as a simplified model of the amplification process in the cell nucleus. PCR with fluorescently labelled dideoxynucleotides commonly employed for DNA sequencing was used to monitor the effect of platinum-based cytostatics on DNA in terms of decrease in labeling efficiency dependent on a presence of the DNA-drug cross-link. Read More

    Electrochemical detection of C-reactive protein using Copper nanoparticles and hybridization chain reaction amplifying signal.
    Anal Biochem 2017 Sep 28;539:1-7. Epub 2017 Sep 28.
    College of Chemistry and Materials Science, The Key Laboratory of Functional Molecular Solids, Ministry of Education, Anhui Key Laboratory of Chem-Biosensing, Anhui Normal University, Wuhu 241000, People's Republic of China. Electronic address:
    In this study, a sandwich-type electrochemical immunosensor for the detection of C-reactive protein (CRP) is described. In design, Copper nanoparticles (Cu NPs) were used for signal tag and hybridization chain reaction (HCR)amplified output signal. The immunosensor fabrication involved three steps: (i) primary antibodies (Ab1) were immobilized on the surface of gold nanoparticles (Au NPs); (ii) the sandwich-type structure formation contained "primary antibodies-antigen-secondary antibodies conjugated with primer (Ab2-S0)"; and (iii) long DNA concatemers intercalating amounts of Cu NPs was linked to the sandwich-type structure via hybridization reaction. Read More

    Recovery and quantification of a myelin oligodendrocyte glycoprotein peptide from rat plasma after protein precipitation.
    Anal Biochem 2017 Sep 27;538:71-73. Epub 2017 Sep 27.
    Department of Chemistry, University of Patras, Rion, 26504, Greece. Electronic address:
    The recovery of high molecular weight peptides from complex biological samples is a challenging task. Herein, a reliable, cost effective and rapid methodology was developed for the recovery and quantification of a myelin oligodendrocyte glycoprotein epitope namely (LysGly)5MOG35-55, from rat plasma. Removal of plasma proteins before quantification of the peptide was achieved after precipitation by an acetonitrile/water/formic acid solution. Read More

    A novel analytical procedure for assaying lysozyme activity using an end-blocked chitotetraose derivative as substrate.
    Anal Biochem 2017 Sep 23;538:64-70. Epub 2017 Sep 23.
    Integrated Bioscience Research Division, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.
    An end-modified β-d-galactosyl chitotetraose derivative [4(4)-O-β-d-galactosyl-β-tri-N-acetylchitotriosyl 2-acetamide-2,3-dideoxy-glucopyranose; Gal(GlcN)3D] was designed and synthesized from chitin tetrasaccharide. The derivative was chemically modified by dehydration of the reducing end GlcN and enzymatic addition of a Gal group to the non-reducing end GlcN. Hydrolysis of Gal(GlcN)3D and related compounds using hen egg-white lysozyme was then examined. Read More

    Glycan profiling of proteins using lectin binding by Surface Plasmon Resonance.
    Anal Biochem 2017 Sep 22;538:53-63. Epub 2017 Sep 22.
    Department of Therapeutic Discovery, Amgen Inc., One Amgen Center Dr., Thousand Oaks, CA 91320, USA. Electronic address:
    Glycan profiling of proteins was studied through their lectin binding activity by Surface Plasmon Resonance (SPR). To validate the method, we monitored specific lectin binding with sequential removal of sugar moieties from human transferrin using specific glycosidases. The results clearly indicated that glycans on the protein can be identified by their selective binding activity to various lectins. Read More

    A compartmentalized culture device for studying the axons of CNS neurons.
    Anal Biochem 2017 Sep 21;539:11-21. Epub 2017 Sep 21.
    Institute of Molecular Medicine, National Tsing Hua University, Hsinchu 30013, Taiwan; Institute of Systems Neuroscience, National Tsing Hua University, Hsinchu 30013, Taiwan. Electronic address:
    We report here the development of a compartmentalized culture device that allows the spatial separation of the somatodendrites and axons of central nervous system (CNS) neurons. The device consists of two compartments separated by a septum constructed by attaching a porous polycarbonate track etch (PCTE) filter on top of a microchannel-filled polydimethylsiloxane (PDMS) membrane. The surface and microchannels of the septum are coated and filled, respectively, with materials that support neuron growth and neurite migration. Read More

    Streamlined purification of fluorescently labeled Escherichia coli phosphate-binding protein (PhoS) suitable for rapid-kinetics applications.
    Anal Biochem 2017 Sep 21. Epub 2017 Sep 21.
    University of Lethbridge, Department of Biological Sciences, 4401 University Dr. W, Lethbridge, AB, T1K 3M4, Canada. Electronic address:
    Fluorescently labeled phosphate-binding proteins can be used as biomolecular tools to measure the release of inorganic phosphate (Pi) from enzymes in real time, enabling the detailed kinetic analysis of dephosphorylating enzymes using rapid-kinetics approaches. Previously reported methods to purify fluorescently labeled phosphate-binding proteins (PhoS) from Escherichia coli are laborious, and a simplified approach is needed. Here, we report the characterization of a cytosol-localized variant (A197C) of PhoS that allows a streamlined purification for subsequent covalent conjugation with a fluorescent dye. Read More

    Menadione-mediated WST1 reduction assay for the determination of metabolic activity of cultured neural cells.
    Anal Biochem 2017 Sep 20;538:42-52. Epub 2017 Sep 20.
    Centre for Biomolecular Interactions Bremen, Faculty 2 (Biology/Chemistry), University of Bremen, Bremen, Germany; Centre for Environmental Research and Sustainable Technology, University of Bremen, Bremen, Germany. Electronic address:
    Cellular reduction of tetrazolium salts to their respective formazans is frequently used to determine the metabolic activity of cultured cells as an indicator of cell viability. For membrane-impermeable tetrazolium salts such as WST1 the application of a membrane-permeable electron cycler is usually required to mediate the transfer of intracellular electrons for extracellular WST1 reduction. Here we demonstrate that in addition to the commonly used electron cycler M-PMS, menadione can also serve as an efficient electron cycler for extracellular WST1 reduction in cultured neural cells. Read More

    Determination of protein thiolation index (PTI) as a biomarker of oxidative stress in human serum.
    Anal Biochem 2017 Sep 20;538:38-41. Epub 2017 Sep 20.
    Department of Life Sciences, Laboratory of Pharmacology and Toxicology, University of Siena, Via A. Moro 2, I-53100 Siena, Italy. Electronic address:
    We have introduced protein thiolation index (PTI), i.e. the molar ratio of the sum of all low molecular mass thiols bound to plasma proteins to protein free cysteinyl residues, as a sensitive biomarker of oxidative stress. Read More

    Colorimetric detection of D-dimer in a paper-based immunodetection device.
    Anal Biochem 2017 Sep 15;538:5-12. Epub 2017 Sep 15.
    iBB- Institute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal. Electronic address:
    A microfluidic paper-based analytical device (μPADs) immunoassay for detection of the blood native biomarker D-dimer is reported. The μPAD is created by wax printing on a single piece of chromatographic paper and combined with an anti-D-dimer capture antibody and conjugates of anti-D-dimer antibody with 40 nm gold nanoparticles. The presence of D-dimer in buffer/simulated plasma samples is successfully reported for concentrations as low as 15 ng D-dimer/mL. Read More

    Ultrasensitive and rapid immuno-detection of human IgE anti-therapeutic horse sera using an electrochemical immunosensor.
    Anal Biochem 2017 Sep 14;538:13-19. Epub 2017 Sep 14.
    FIOCRUZ, Center for Technological Development in Health (CDTS)/National Institute of Science and Technology on Innovation in Neglected Population Diseases (INCT-IDPN) FIOCRUZ, Rio de Janeiro, RJ, Brazil; FIOCRUZ, Oswaldo Cruz Institute, Laboratory of Experimental and Computational Biochemistry of Pharmaceuticals, Rio de Janeiro, RJ, Brazil; Federal Fluminense University, Biology Institute, Cellular and Molecular Department, Niterói, Rio de Janeiro, Brazil. Electronic address:
    Antivenom allergy disease mediated by patient IgE is an important public health care concern. To improve detection of hypersensitive individuals prior to passive antibody therapy, an amperometric immunosensor was developed to detect reactive human IgE. Whole horse IgG3 (hoIgG3) was immobilized onto the surface of carbon or gold screen-printed electrodes through a cross-linking solution of glutaraldehyde on a chitosan film. Read More

    Ultrasensitive microRNA-21 detection based on DNA hybridization chain reaction and SYBR Green dye.
    Anal Biochem 2017 Sep 15;538:20-25. Epub 2017 Sep 15.
    College of Chemistry and Material Science, Shandong Agricultural University, 271018, Taian, Shandong, PR China.
    It is extremely important for quantifying trace microRNAs in the biomedical applications. In this study, an ultrasensitive, rapid and efficient label-free fluorescence method was proposed and applied for detecting microRNA-21 in serum of gastric cancer patients based on DNA hybridization chain reaction (HCR). DNA H1 and DNA H2 were designed and used as hairpin probes, the HCR was proceeded in the presence of target microRNAs. Read More

    Fluorescence and magnetic nanocomposite Fe3O4@SiO2@Au MNPs as peroxidase mimetics for glucose detection.
    Anal Biochem 2017 Sep 14;538:26-33. Epub 2017 Sep 14.
    College of Science, Sichuan Agricultural University, Ya'an 625014, China. Electronic address:
    In this paper, multifunction nanoparticles (MNPs), Fe3O4@SiO2@Au MNPs, with properties of superparamagnetism, fluorescence and peroxidase-like catalytic activity were synthesized in the aqueous phase. The synthesized composites were characterized by scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), X-ray diffraction (XRD), Fourier translation infrared spectrum (FT-IR) and fluorometer. The results show that the multifunctional nanomaterials have good magnetic and fluorescence properties. Read More

    A novel electrochemical immunosensor based on ITO modified by carboxyl-ended silane agent for ultrasensitive detection of MAGE-1 in human serum.
    Anal Biochem 2017 Sep 12;537:84-92. Epub 2017 Sep 12.
    Çanakkale Onsekiz Mart University, Faculty of Engineering, Bioengineering Department, Çanakkale, Turkey.
    A new, low-cost electrochemical immunosensor was developed for rapid detection of Melanoma-associated antigen 1 (MAGE-1), a cancer biomarker. The fabrication procedure of immunosensor was based on the covalent immobilization of anti-MAGE-1, biorecognition molecule, on ITO electrode by carboxyethylsilanetriol (CTES) monolayer. The biosensing MAGE-1 antigen was monitored by using electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) technique. Read More

    A multiplex protein-free lateral flow assay for detection of microRNAs based on unmodified molecular beacons.
    Anal Biochem 2017 Sep 11;537:99-105. Epub 2017 Sep 11.
    Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
    Lateral flow assays (LFAs) have promising potentials for point-of-care applications. Recently, many LFAs have been reported that are based on hybridization of oligonucleotide strands. Mostly, biotinylated capture DNAs are immobilized on the surface of a nitrocellulose membrane via streptavidin interactions. Read More

    Genotyping of single nucleotide polymorphism by probe-gated silica nanoparticles.
    Anal Biochem 2017 Sep 8;537:78-83. Epub 2017 Sep 8.
    Yeditepe University, School of Medicine, Department of Biophysics, Istanbul, Turkey. Electronic address:
    The development of simple, reliable, and rapid approaches for molecular detection of common mutations is important for prevention and early diagnosis of genetic diseases, including Thalessemia. Oligonucleotide-gated mesoporous nanoparticles-based analysis is a new platform for mutation detection that has the advantages of sensitivity, rapidity, accuracy, and convenience. A specific mutation in β-thalassemia, one of the most prevalent inherited diseases in several countries, was used as model disease in this study. Read More

    Gas chromatography tandem mass spectrometry offers advantages for urinary steroids analysis.
    Anal Biochem 2017 Sep 6;538:34-37. Epub 2017 Sep 6.
    University/British Heart Foundation Centre for Cardiovascular Science, Queen's Medical Research Institute, University of Edinburgh, 47, Little France Crescent, Edinburgh, EH16 2TJ, UK. Electronic address:
    Gas chromatography mass spectrometry has been the lynchpin of clinical assessment of steroid profiles for ∼3 decades. The improvements in assay performance offered by tandem mass spectrometry were assessed. Across the spectrum of glucocorticoid and androgen analytes tested, limits of detection and quantitation were ∼20 fold lower with triple than single quadrupole systems, but the more noticeable improvement was that signal to noise was substantially improved and the linear range wider. Read More

    Adsorption effects of the doping relevant peptides Insulin Lispro, Synachten, TB-500 and GHRP 5.
    Anal Biochem 2017 Sep 5;537:69-71. Epub 2017 Sep 5.
    Ghent University, Department of Clinical Chemistry, Microbiology and Immunology, Doping Control Laboratory, Technologiepark 30 B, B-9052 Zwijnaarde, Belgium.
    The tendency of peptides to adsorb to surfaces can raise a concern in variety of analytical fields where the qualitative/quantitative measurement of low concentration analytes (ng/mL-pg/mL) is required. To demonstrate the importance of using the optimal glassware/plasticware, four doping relevant model peptides (GHRP 5, TB-500, Insulin Lispro, Synachten) were chosen and their recovery from various surfaces were evaluated. Our experiments showed that choosing expensive consumables with low-bind characteristics is not beneficial in all cases. Read More

    Method to minimize ozone effect on Cy5 fluorescent intensity in DNA microarrays.
    Anal Biochem 2017 Sep 4;538:1-4. Epub 2017 Sep 4.
    Department of Obstetrics and Gynecology, Cheil General Hospital and Women's Healthcare Center, Dankook University College of Medicine, 17, Seoae ro 1 Gil, Jung gu, Seoul, Republic of Korea. Electronic address:
    Cyanine 5 (Cy5) is an established fluorescent dye in microarray analysis. It is degraded rapidly when exposed to atmospheric ozone during post-hybridization washes, which leads to loss of fluorescent intensity. To minimize this undesirable effect, we coated microarray slides with sodium dodecyl sulfate (SDS) solution at post-hybridization washes. Read More

    A novel method of multiple nucleic acid detection: Real-time RT-PCR coupled with probe-melting curve analysis.
    Anal Biochem 2017 Sep 4;537:50-55. Epub 2017 Sep 4.
    School of Life Science and Biopharmaceutics, Shenyang Pharmaceutical University, Shenyang 110016, PR China. Electronic address:
    A novel method, real-time reverse transcription PCR (real-time RT-PCR) coupled with probe-melting curve analysis, has been established to detect two kinds of samples within one fluorescence channel. Besides a conventional TaqMan probe, this method employs another specially designed melting-probe with a 5' terminus modification which meets the same label with the same fluorescent group. By using an asymmetric PCR method, the melting-probe is able to detect an extra sample in the melting stage effectively while it almost has little influence on the amplification detection. Read More

    An improved amperometric creatinine biosensor based on nanoparticles of creatininase, creatinase and sarcosine oxidase.
    Anal Biochem 2017 Sep 1;537:41-49. Epub 2017 Sep 1.
    Department of Biochemistry, M.D. University, Rohtak 124001 India. Electronic address:
    An improved amperometric biosensor for detection of creatinine was developed based on immobilization of nanoparticles (NPs) of creatininase (CA), creatinase (CI), and sarcosine oxidase (SOx) onto glassy carbon (GC) electrode. Transmission electron microscopy (TEM) and fourier transform infrared spectroscopy (FTIR) were employed for characterization of enzyme nanoparticles (ENPs). The GC electrode was characterized by scanning electron microscopy (SEM), cyclic voltammetry (CV) and electrochemical impedance spectra (EIS) at different stages of its amendment. Read More

    Chemiluminescence immunoassays for estradiol and ethinylestradiol based on new biotinylated estrogen derivatives.
    Anal Biochem 2017 Sep 7;537:63-68. Epub 2017 Sep 7.
    Univ. Perpignan Via Domitia, Biocapteurs-Analyses-Environnement, 66860, Perpignan, France; Laboratoire de Biodiversité et Biotechnologies Microbiennes, USR 3579 Sorbonne Universités (UPMC) Paris 6 et CNRS Observatoire Océanologique, 66650, Banyuls-sur-Mer, France. Electronic address:
    New chemiluminescence-based immunoassays for sensitive detection of 17-β estradiol (E2) and ethinylestradiol (EE2) are described on the basis of the use of biotinylated estrogen derivatives. Estrogen derivatives bearing a carboxylic group (E2-COOH and EE2-COOH) on C-3 position were synthesized, covalently bound to aminated biotin and subsequently immobilized on avidin-coated microtiter plates. The assay principle was based on competition between free and immobilized estrogens for their binding to primary antibodies, with subsequent revelation using horseradish peroxidase (HRP)-labeled secondary antibodies. Read More

    A miniaturized assay for kinetic characterization of the Na(+)-translocating NADH:ubiquinone oxidoreductase from Vibrio cholerae.
    Anal Biochem 2017 Sep 7;537:56-59. Epub 2017 Sep 7.
    Institute of Microbiology, University of Hohenheim (Stuttgart), Garbenstraße 30, 70599 Stuttgart, Germany. Electronic address:
    We demonstrate the miniaturization of an enzymatic assay for the determination of NADH oxidation and quinone reduction by the Na(+) -translocating NADH quinone oxidoreductase (NQR) in the 96-well plate format. The assay is based on the spectrophotometric detection of NADH consumption and quinol formation. We validated the new method with known inhibitors of the NQR and optimized conditions for high-throughput screening as demonstrated by excellent Z-factors well above the accepted threshold (≥0. Read More

    A generic approach for simultaneous measurements of total antibody and cleavable antibody-conjugated drug by LC/MS/MS.
    Anal Biochem 2017 Sep 1;537:33-36. Epub 2017 Sep 1.
    Millennium Pharmaceuticals, Inc., A Wholly Owned Subsidiary of Takeda Pharmaceutical Company Limited, 35 Landsdowne Street, Cambridge, MA 02139, USA. Electronic address:
    The current industry practice for antibody-drug conjugate (ADC) bioanalysis includes quantification of total antibody and antibody-conjugated drug. Here, we report a novel 2-in-1 approach for measuring total antibody and protease-cleavable conjugated drug Monomethyl Auristatin E (MMAE) concurrently. This allows for the determination of the DAR (Drug Antibody Ratio) for in vivo samples, with a 3-orders linear range based on total antibody concentration from 0. Read More

    Continuous liquid feeding: New method to study pesticides toxicity in Drosophila melanogaster.
    Anal Biochem 2017 Sep 1;537:60-62. Epub 2017 Sep 1.
    Laboratório de Estudos Físico-Químicos e Produtos Naturais (LEFQPN), Universidade Federal do Pampa, Uruguaiana, RS, Brazil. Electronic address:
    Fly fruit Drosophila melanogaster (DM) has been extensively employed as an in vivo model system to study pesticides toxicity. Pesticide administration to the fly traditionally involves feeding in an agar-gelled feed fly's medium (AM). However, AM method has several limitations such as uncertainty regarding the bioavailability and amount of pesticides ingested. Read More

    IEF peptide fractionation method combined to shotgun proteomics enhances the exploration of rice milk proteome.
    Anal Biochem 2017 Aug 30;537:72-77. Epub 2017 Aug 30.
    Department of Biotechnology, Proteomics and Mass Spectrometry Lab, University of Verona, Strada le Grazie 15, 37134, Verona, Italy.
    We conducted a proteomics study in order to detect the proteomic method which provides the most complete characterization of the proteins of rice milk. In particular, we compared the results obtained from LC-MS/MS after protein precipitation with acetone or TCA, as well as the results obtained from LC-MS/MS after protein prefractionation based on SDS-PAGE (GeLC-MS/MS) or ProteoMiner™ technology (ProteoMiner-LC-MS/MS), and after peptide prefractionation based on IEF (pIEF-LC-MS/MS). A total of 158 protein species have been detect in rice milk. Read More

    Preparation of wool follicles for proteomic studies.
    Anal Biochem 2017 Aug 30;539:8-10. Epub 2017 Aug 30.
    AgResearch Ltd, Lincoln Research Centre, Christchurch 8140, New Zealand.
    A variety of techniques were applied to wool follicles stored in William's E culture medium to optimise the extraction of keratin and keratin associated proteins (KAPs). A time course study indicated that the maximum storage time for live skin in this buffer at 20 °C was 24 h, after which degradative loss of protein became significant. Maceration of the skin for 10 min followed by reciprocal action shaking for 14 h had a detrimental effect on keratin extractability. Read More

    Isotope-dilution liquid chromatography-tandem mass spectrometry for sensitive quantification of human insulin in serum using derivatization-technique.
    Anal Biochem 2017 Aug 30;537:26-32. Epub 2017 Aug 30.
    Bio-medical Standards Group, Research Institute for Material and Chemical Measurement, National Metrology Institute of Japan (NMIJ), National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba C-3, Ibaraki 305-8563, Japan.
    An isotope-dilution mass spectrometry (IDMS) method for measuring insulin levels in human serum was developed using C-terminal-derivatization method coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). The carboxyl groups of Glu-C-cleavage products were derivatized with 1-(2-pyrimidinyl)piperazine to increase MS/MS sensitivity and IDMS quantification, resulting in increases in LC-MS/MS peak areas of derivatized Glu-C-cleavage products of human insulin by ∼23-(A5-17 peptide) to 49-fold(B14-21 peptide), respectively, as compared with results observed in the absence of derivatization. Separation was achieved on a C18 column by gradient elution at 0. Read More

    Capillary electrophoresis - Mass spectrometry metabolomics analysis revealed enrichment of hypotaurine in rat glioma tissues.
    Anal Biochem 2017 Aug 25;537:1-7. Epub 2017 Aug 25.
    Department of Surgery, Dalian Sixth People's Hospital, Dalian, 116031, China. Electronic address:
    Glioma is one of the most lethal brain malignancies with unknown etiologies. Many metabolomics analysis aiming at diverse kinds of samples had been performed. Due to the varied adopted analytical platforms, the reported disease-related metabolites were not consistent across different studies. Read More

    A fluorescence method for determination of glucose transport by intestinal BBMV of common carp.
    Anal Biochem 2017 Aug 26;537:20-25. Epub 2017 Aug 26.
    College of Fisheries, Henan Normal University, Xinxiang 453007, China. Electronic address:
    Epithelial brush-border membrane vesicles (BBMVs) were isolated from the intestine of common carp and studied systematically by enzyme activity, transmission electron microscopy and immunoblotting. The uptake time course and the substrate concentration effect were assessed, and then, the ability of phlorizin and cytochalasin B to inhibit uptake was analyzed. The results show that sucrase, alkaline phosphatase and Na(+)-K(+)-ATPase activities in these vesicles were enriched 7. Read More

    Imaged capillary isoelectric focusing in native condition: A novel and successful example.
    Anal Biochem 2017 Aug 24;537:13-19. Epub 2017 Aug 24.
    Department of Biologics Process Development, Bristol-Myers Squibb, USA.
    Imaged capillary isoelectric focusing (icIEF) separates ampholytic components of biomolecules in an electric field according to their isoelectric points and has been used for protein charge variants quantification and characterization. Denaturants are ordinarily incorporated into icIEF to stabilize charge species in solution. In certain circumstances, however, denaturants are detrimental to stable isoelectric separation of proteins due to their unique structural and biophysical features, such as an aggregation-prone antibody we encountered recently. Read More

    Measurement of O-GlcNAcylated endothelial nitric oxide synthase by using 2',5'-ADP-Sepharose pull-down assay.
    Anal Biochem 2017 Aug 24;537:8-12. Epub 2017 Aug 24.
    Division of Cardiology, The First Affiliated Hospital, Chongqing Medical University, Chongqing, 400016, China; Institute of Life Science, Chongqing Medical University, Chongqing, 400010, China; Davis Heart and Lung Research Institute, Division of Cardiovascular Medicine, Department of Molecular and Cellular Biochemistry, The Ohio State University College of Medicine, 473 West 12th Avenue, Columbus, OH, 43210, USA. Electronic address:
    Endothelial nitric oxide synthase (eNOS) plays central roles in cardiovascular regulation and disease. eNOS function is critically affected by O-linked N-acetylglucosamine (O-GlcNAc) modification. The present method for measuring O-GlcNAcylated eNOS relies on immunoprecipitation. Read More

    NADPH oxidase activity: Spectrophotometric determination of superoxide using pyrogallol red.
    Anal Biochem 2017 Aug 24;536:96-100. Epub 2017 Aug 24.
    Department of Pharmacy, Faculty of Chemistry, Pontificia Universidad Católica de Chile, Chile. Electronic address:
    A simple and fast spectrophotometric methodology able to quantify superoxide released by NADPH oxidase from differentiated promyelocytic leukaemia (HL-60) cells using pyrogallol red is described.The latter is based on the known stoichiometry of the reaction between superoxide and pyrogallol red and the inability of pyrogallol red to react with hydrogen peroxide. In addition, we developed a 96-wells microplate-based method able to determine NADPH oxidase activity. Read More

    Expression and purification of functional insulin and insulin-like growth factor 1 holoreceptors from mammalian cells.
    Anal Biochem 2017 Aug 19;536:69-77. Epub 2017 Aug 19.
    Department of Physiology and Biophysics, Stony Brook University, NY, United States; Department of Veterans Affairs Medical Center, Northport, NY, United States. Electronic address:
    The insulin receptor (IR) and insulin-like growth factor 1 receptor (IGF1R) are receptor tyrosine kinases (RTKs) involved in the regulation of many important cellular processes. The current proposed models of activation are derived from structural studies using soluble extracellular domains and cytoplasmic tyrosine kinase domains. Preparations of full length IR and IGF1R have been hampered by the need for unconventional affinity chromatography resins and/or harsh eluting conditions. Read More

    New method for estimating clustering of DNA lesions induced by physical/chemical mutagens using fluorescence anisotropy.
    Anal Biochem 2017 Aug 18;536:78-89. Epub 2017 Aug 18.
    Radiation Biochemistry and Biological Function, Research Reactor Institute, Kyoto University, Kumatori, Sennan, Osaka 590-0494, Japan.
    We have developed a new method for estimating the localization of DNA damage such as apurinic/apyrimidinic sites (APs) on DNA using fluorescence anisotropy. This method is aimed at characterizing clustered DNA damage produced by DNA-damaging agents such as ionizing radiation and genotoxic chemicals. A fluorescent probe with an aminooxy group (AlexaFluor488) was used to label APs. Read More

    Methylene blue as a lignin surrogate in manganese peroxidase reaction systems.
    Anal Biochem 2017 Aug 18;537:37-40. Epub 2017 Aug 18.
    School of Chemical, Biological, and Environmental Engineering, 116 Johnson Hall, Oregon State University, Corvallis, OR 97331, USA. Electronic address:
    Manganese peroxidase (MnP) is associated with lignin degradation and is thus relevant to lignocellulosic-utilization technologies. Technological applications require reaction mixture optimization. A surrogate substrate can facilitate this if its susceptibility to degradation is easily monitored and mirrors that of lignin. Read More

    The role of human monoacylglycerol lipase (hMAGL) binding pocket in breakup of unsaturated phospholipid membranes.
    Anal Biochem 2017 Aug 17;536:90-95. Epub 2017 Aug 17.
    Center for Drug Discovery, Northeastern University, Boston, MA, 02115, United States; Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA, 02115, United States; Pharmaceutical Sciences Department, Northeastern University, Boston, MA, 02115, United States.
    Human monoacylglycerol lipase (hMAGL) plays a key role in homeostatic tuning of the endocannabinoid signaling system and supports aggressive tumorogenesis, making this enzyme a promising therapeutic target. hMAGL features a membrane-associated lid domain that regulates entry of endocannabinoid lipid substrates into the hydrophobic channel accessing the active site, likely from the membrane bilayer. The present work applied simultaneous surface plasmon resonance and electrochemical impedance spectroscopy measurements to show that, in absence of the substrate, hMAGL can remove phospholipid molecules from the membrane and, thereby, disintegrate pre-formed, intact, tethered phospholipid bilayer membrane mimetics (tBLMs) composed of unsaturated phosphatidylcholines. Read More

    TEMPORARY REMOVAL: Enhanced post wash retention of combed DNA molecules by varying multiple combing parameters.
    Anal Biochem 2017 Aug 15;536:45-50. Epub 2017 Aug 15.
    Birla Institute of Scientific Research, Jaipur, India. Electronic address:
    The publisher regrets that this article has been temporarily removed. A replacement will appear as soon as possible in which the reason for the removal of the article will be specified, or the article will be reinstated. The full Elsevier Policy on Article Withdrawal can be found at https://www. Read More

    DNA aptamer identification and characterization for E. coli O157 detection using cell based SELEX method.
    Anal Biochem 2017 Aug 14;536:36-44. Epub 2017 Aug 14.
    Department of Molecular Biology, Pasteur Institute of Iran, Pasteur Ave., Tehran 13164, Iran.
    Escherichia coli (E. coli) O157:H7 is a foodborne pathogen that causes symptoms in humans. Its rapid identification should be considered to avoid toxic effects of the pathogen. Read More

    Real-time qRT-PCR assay for the detection of miRNAs using bi-directional extension sequences.
    Anal Biochem 2017 Aug 12;536:32-35. Epub 2017 Aug 12.
    Department of Pharmaceutical Engineering, Soonchunhyang University, 22 Soonchunhyang-ro, Shinchang-myeon, Asan-si, Chungcheongnam-do 31538, Republic of Korea. Electronic address:
    Highly specific detection of miRNAs was performed using a novel bi-directional extension (BDE) assay. After reverse transcription, the cDNA was hybridized to a uniquely designed specific BDE sequence; this cDNA-BDE hybrid forms the PCR template. The PCR template was amplified in a SYBR Green-based quantitative real-time PCR. Read More

    A miniaturized peptidyl-prolyl isomerase enzyme assay.
    Anal Biochem 2017 Aug 11;536:59-68. Epub 2017 Aug 11.
    College of Life and Environmental Sciences, University of Exeter, Exeter EX4 4QD, United Kingdom. Electronic address:
    Prolyl-peptidyl isomerases (PPIases) are enzymes that are found in all living organisms. They form an essential part of the cellular protein folding homeostasis machinery. PPIases are associated with many important human diseases, e. Read More

    A method for extracting and characterizing RNA from urine: For downstream PCR and RNAseq analysis.
    Anal Biochem 2017 Aug 10;536:8-15. Epub 2017 Aug 10.
    University of Minnesota Medical School - Duluth, 1035 University Drive, Duluth, MN 55812-3031, USA. Electronic address:
    Readily accessible samples such as urine or blood are seemingly ideal for differentiating and stratifying patients; however, it has proven a daunting task to identify reliable biomarkers in such samples. Noncoding RNA holds great promise as a source of biomarkers distinguishing physiologic wellbeing or illness. Current methods to isolate and characterize RNA molecules in urine are limited. Read More

    Designing binding kinetic assay on the bio-layer interferometry (BLI) biosensor to characterize antibody-antigen interactions.
    Anal Biochem 2017 Aug 10;536:16-31. Epub 2017 Aug 10.
    Biomolecular HTS Center, Therapeutic Proteins, Regeneron Pharmaceuticals, 777 Old Saw Mill River Road, Tarrytown, NY 10591, USA.
    The Octet biosensors provide a high-throughput alternative to the well-established surface plasmon resonance (SPR) and SPR imaging (SPRi) biosensors to characterize antibody-antigen interactions. However, the utility of the Octet biosensors for accurate and reproducible measurement of binding rate constants of monoclonal antibodies (mAbs) is limited due to challenges such as analyte rebinding, and mass transport limitation (MTL). This study focuses on addressing these challenges and provides experimental conditions to reliably measure kinetics of mAb-antigen interactions. Read More

    Graphene oxide layer decorated gold nanoparticles based immunosensor for the detection of prostate cancer risk factor.
    Anal Biochem 2017 Aug 5;536:51-58. Epub 2017 Aug 5.
    Analytical Chemistry Group, Chemical Sciences and Technology Division, CSIR-North East Institute of Science & Technology, Academy of Scientific and Innovative Research, Jorhat, 785006, Assam, India. Electronic address:
    In this work, we report a novel electrochemical immunosensor based on gold nanoparticles deposited on the surface of graphene oxide layers and was used for immobilization of monoclonal anti-PSA antibody via EDC/NHS coupling method to detect prostate-specific antigen (PSA), a valuable biomarker for early detection of prostate cancer. To confirm the functionality of the antibody, we performed immunofluorescence staining using human prostate adenocarcinoma cells, LNCaP. Scanning Electron Microscopy (SEM), cyclic voltammetry, and other electrochemical techniques were used to characterize the resulting electrode surface. Read More

    Neutral Red versus MTT assay of cell viability in the presence of copper compounds.
    Anal Biochem 2017 Oct 2;535:43-46. Epub 2017 Aug 2.
    Département de Chimie and Centre de Recherche BioMed, Université du Québec à Montréal, C.P. 8888, Succ. Centre-ville, Montreal, Quebec H3C 3P8, Canada. Electronic address:
    Copper is essential for numerous physiological functions, and copper compounds may display therapeutic as well as cytotoxic effects. The MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) assay is a standard test largely used in cytotoxicity studies. This report shows that low micromolar levels of copper compounds such as Cu(II)Urea2, Cu(II)Ser2 and CuCl2 can interfere with the MTT assay making improper the detection of formazan product of MTT reduction. Read More

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