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    26440 results match your criteria Analytical Biochemistry [Journal]

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    Development of a borreliosis assay: Mannan coated polyethylene sinter bodies as a new platform technology.
    Anal Biochem 2017 Dec 6;543:55-61. Epub 2017 Dec 6.
    Institute of Pharmaceutical Chemistry, Philipps-Universität Marburg, Marbacher Weg 6, 35032 Marburg, Germany.
    Rapid diagnosis of Lyme borreliosis has been carried out on chemically modified porous polyethylene sinter bodies. Photografting of 2-propenol on sinter body's surface was performed as a first step, introducing active hydroxyl groups as a result of polyalcohol formation. The hydroxyl groups were used for further immobilization and could be linked via 3-aminopropyltriethoxysilane (APTES) to polysaccharides like mannan. Read More

    Rapid profiling method for mammalian feces short chain fatty acids by GC-MS.
    Anal Biochem 2017 Dec 6;543:51-54. Epub 2017 Dec 6.
    Anicom Specialty Medical Institute Inc., 8-17-1, Nishi Shinjuku, Shinjuku-ku, Tokyo, Japan.
    Short chain fatty acids (SCFAs) are key feces metabolites generated by gut bacteria fermentation. Despite the importance of profiling feces SCFAs, technical difficulties in analysis remain due to their volatility and hydrophilicity. We improve previous protocols to profile SCFAs and optimize the metabolite profiling platform for mammalian feces samples. Read More

    Microplate chemiluminescent assay for HBV DNA detection using 3-(10'-phenothiazinyl)propionic acid/N-morpholinopyridine pair as enhancer of HRP-catalyzed chemiluminescence.
    Anal Biochem 2017 Dec 5;543:33-36. Epub 2017 Dec 5.
    Department of Chemistry, Lomonosov Moscow State University, Leninskie gory, Moscow 119991, Russia. Electronic address:
    A sensitive sandwich assay for hepatitis B virus (HBV) DNA detection based on use of commercial CL-ELISA microplates was developed. To reveal the target the covalent conjugate of reporter oligonucleotide and horseradish peroxidase (HRP) was synthesized. An employment of enhanced chemiluminescence reaction, where 3-(10'-phenothiazinyl)propionic acid/N-morpholinopyridine pair was used as enhancer of HRP-catalyzed chemiluminescence, permitted to measure the enzyme activity of the conjugate with high sensitivity. Read More

    Exploring three different expression systems for recombinant expression of globins: Escherichia coli, Pichia pastoris and Spodoptera frugiperda.
    Anal Biochem 2017 Dec 5;543:62-70. Epub 2017 Dec 5.
    Department of Biomedical Sciences, University of Antwerp, Universiteitsplein 1, Antwerp 2610, Belgium. Electronic address:
    Globins are among the best investigated proteins in biological and medical sciences and represent a prime tool for the study of the evolution of genes and the structure-function relationship of proteins. Here, we explore the recombinant expression of globins in three different expression systems: Escherichia coli, Pichia pastoris and the baculovirus infected Spodoptera frugiperda. We expressed two different human globin types in these three expression systems: I) the well-characterized neuroglobin and II) the uncharacterized, circular permutated globin domain of the large chimeric globin androglobin. Read More

    Proteomic profiling of large myofibrillar proteins from dried and long-term stored polyacrylamide gels.
    Anal Biochem 2017 Dec 2;543:8-11. Epub 2017 Dec 2.
    Department of Biology, Maynooth University, National University of Ireland, Maynooth, Ireland. Electronic address:
    A method for the utilization of dried polyacrylamide gels from the pre-proteomic era is described in order to enable the mass spectrometric analysis of long-term stored protein preparations. The in-gel digestion of high-molecular-mass proteins embedded in a 20-year old gel was carried out following gel re-swelling and resulted in the proteomic identification of a large number of proteins, including 3400 kDa titin, 800 kDa nebulin and myosin heavy chains of 220 kDa from rabbit skeletal muscle. These findings demonstrate that dried protein gels from past biochemical analyses can be successfully reused and analyzed by modern and refined mass spectrometric techniques. Read More

    Voltammetric determination of thujone in herbal matrices in the presence of Triton X-100.
    Anal Biochem 2017 Dec 1;543:12-20. Epub 2017 Dec 1.
    AGH University of Science and Technology, Faculty of Materials Science and Ceramics, Mickiewicza 30, 30-059 Kraków, Poland. Electronic address:
    A simple, sensitive and rapid method of low concentration of thujone determination using Controlled Growth Mercury Drop Electrode and Differential Pulse Voltammetry was described. The reduction of thujone in phosphoric acid or potassium nitrate, involves a quasi-reversible and adsorption-controlled two-electron process. In voltammetric experiments the addition of ethanol and Triton X-100 is recommended. Read More

    Design and application of a fluorogenic assay for monitoring inflammatory caspase activity.
    Anal Biochem 2017 Dec 1;543:1-7. Epub 2017 Dec 1.
    DePaul University, Department of Chemistry, 1110 W. Belden Ave., Chicago, IL 60614, USA. Electronic address:
    Various fluorogenic assays exist for monitoring the activity of inflammatory caspases. However, there are no continuous assays that provide C-terminal substrate sequence specificity for inflammatory caspases. As a first step towards this, we have developed a continuous in vitro assay that relies on monitoring emission from tryptophan after cleavage of a quenching coumarin chromophore. Read More

    Use of alternative alkali chlorides in RT and PCR of polynucleotides containing G quadruplex structures.
    Anal Biochem 2017 Dec 2;543:43-50. Epub 2017 Dec 2.
    Centro de Investigación en Ciencia Aplicada y Tecnología Avanzada, Instituto Politécnico Nacional, Cerro Blanco 141, Colonia Colinas del Cimatario, Querétaro, QRO 76090, Mexico. Electronic address:
    Several alkali chlorides were compared for their use in reverse transcription (RT) and PCR of different types of nucleic acid templates. On a test region of biological DNA incapable of forming G quadruplex (G4) structures, Taq DNA polymerase showed similar PCR performance with 50 mM KCl, CsCl, LiCl, and NaCl. In contrast, on a synthetic model polydeoxyribonucleotide prone to G4 formation, good PCR amplification was obtained with 50 mM CsCl, but little or none with LiCl or KCl. Read More

    ImageJ-based semiautomatic method to analyze senescence in cell culture.
    Anal Biochem 2017 Nov 28;543:30-32. Epub 2017 Nov 28.
    Aragon Institute of Health Sciences & IIS Aragon, 50009, Zaragoza, Spain.
    β-Galactosidase accumulates in the lysosomes of senescent cells of certain tissues. Cell staining with X-gal is a common procedure to detect senescent cells in culture. However, the organelle nature of the staining makes automatic count impossible, requiring time-consuming manual counting or expensive alternative techniques such as flow cytometry to effectively determine the amount of stained cells. Read More

    Hydrophilic modified magnetic multi-walled carbon nanotube for dispersive solid/liquid phase microextraction of sunitinib in human samples.
    Anal Biochem 2017 Nov 29;542:76-83. Epub 2017 Nov 29.
    Department of Chemistry, Payame Noor University, Tehran, P.O. Box 19395-4697, Iran. Electronic address:
    In this paper, a novel approach for the efficient microextraction and determination of anticancer drug, sunitinib from human samples is described. We synthesized a new nanocomposite; honey coated magnetic multi-walled carbon nanotubes (Honey@magnetic-CNTs). This nanocomposite retains the magnetic properties of individual magnetic nanoparticles (MNPs) and can be effectively separated under an external magnetic field. Read More

    Contact lens to measure individual ion concentrations in tears and applications to dry eye disease.
    Anal Biochem 2017 Nov 26;542:84-94. Epub 2017 Nov 26.
    Center for Fluorescence Spectroscopy, Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, 725 W. Lombard Street, Baltimore, MD 21201, USA.
    Dry eye disease (DED) affects millions of individuals in the United States and worldwide, and the incidence is increasing with an aging population. There is widespread agreement that the measurement of total tear osmolarity is the most reliable test, but this procedure provides only the total ionic strength and does not provide the concentration of each ionic species in tears. Here, we describe an approach to determine the individual ion concentrations in tears using modern silicone hydrogel (SiHG) contact lenses. Read More

    A high-throughput screening campaign to identify inhibitors of DXP reductoisomerase (IspC) and MEP cytidylyltransferase (IspD).
    Anal Biochem 2017 Nov 24;542:63-75. Epub 2017 Nov 24.
    Department of Chemistry and Biochemistry, George Mason University, Manassas, VA 20110, USA. Electronic address:
    The rise of antibacterial resistance among human pathogens represents a problem that could change the landscape of healthcare unless new antibiotics are developed. The methyl erythritol phosphate (MEP) pathway represents an attractive series of targets for novel antibiotic design, considering each enzyme of the pathway is both essential and has no human homologs. Here we describe a pilot scale high-throughput screening (HTS) campaign against the first and second committed steps in the pathway, catalyzed by DXP reductoisomerase (IspC) and MEP cytidylyltransferase (IspD), using compounds present in the commercially available LOPAC1280 library as well as in an in-house natural product extract library. Read More

    New spectrophotometric assay for assessments of catalase activity in biological samples.
    Anal Biochem 2017 Nov 22;542:29-33. Epub 2017 Nov 22.
    Chemistry Dept., Faculty of Education for women, Kufa University, Najaf city, Najaf Governorate, Iraq. Electronic address:
    A novel, simple, and accurate colorimetric assay was established for assessments of catalase activity in biological fluids and tissues. H2O2 dissociation rates are directly proportional to catalase activity, and the principle of the present assay is based on reactions of ammonium metavanadate with H2O2 under acidic conditions. The resulting reduction of vanadium (V) to vanadium (III) produces a red-orange peroxovanadium complex with absorbance maxima at 452 nm. Read More

    Polymerization of hexamethylene diisocyanate in solution and a 260.23 m/z [M+H]+ ion in exposed human cells.
    Anal Biochem 2017 Nov 24;543:21-29. Epub 2017 Nov 24.
    Department of Internal Medicine, Yale University, New Haven, CT 06520, United States.
    Hexamethylene diisocyanate (HDI) is an important industrial chemical that can cause asthma, however pathogenic mechanisms remain unclear. Upon entry into the respiratory tract, HDI's N=C=O groups may undergo nucleophilic addition (conjugate) to host molecules (e.g. Read More

    Evaluation of digestion methods for analysis of trace metals in mammalian tissues and NIST 1577c.
    Anal Biochem 2017 Nov 24;543:37-42. Epub 2017 Nov 24.
    University of South Florida, Tampa, Chemistry Department & Florida Center for Drug Discovery & Innovation (CDDI), USA. Electronic address:
    Digestion techniques for ICP analysis have been poorly studied for biological samples. This report describes an optimized method for analysis of trace metals that can be used across a variety of sample types. Digestion methods were tested and optimized with the analysis of trace metals in cancerous as compared to normal tissue as the end goal. Read More

    15N CEST data and traditional model-free analysis capture fast internal dynamics of DJ-1.
    Anal Biochem 2017 Nov 21;542:24-28. Epub 2017 Nov 21.
    Department of Chemistry, University of Nebraska-Lincoln, Lincoln, NE 68588-0304, USA; Nebraska Center for Integrated Biomolecular Communication, University of Nebraska-Lincoln, Lincoln, NE 68588-0304, USA. Electronic address:
    Previous studies have shown that relaxation parameters and fast protein dynamics can be quickly elucidated from 15N-CEST experiments [1]. Longitudinal R1 and transverse R2 values were reliably derived from fitting of CEST profiles. Herein we show that 15N-CEST experiments and traditional modelfree analysis provide the internal dynamics of three states of human protein DJ-1 at physiological temperature. Read More

    One-step FPLC-size-exclusion chromatography procedure for purification of rDMBT1 6 kb with increased biological activity.
    Anal Biochem 2017 Nov 21;542:16-19. Epub 2017 Nov 21.
    Lundbeckfonden Center of Excellence NanoCAN, University of Southern Denmark, Odense, Denmark; Department of Cancer and Inflammation Research, Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark; Department of Oncology, Odense University Hospital, Odense, Denmark. Electronic address:
    Deleted in Malignant Brain Tumor 1 (DMBT1, alias SAG or gp340) is a pattern recognition receptor involved in immune defense, cell polarization, differentiation and regeneration. To investigate the role of the protein in physiological and pathological processes, the protein has often been isolated from saliva or produced in vitro and purified by a multistep affinity purification procedure using bacteria, followed by FPLC. Here, we compared a simple, one-step FPLC-SEC protocol for purification of recombinant DMBT1 6 kb, with that of the standard bacteria affinity purification-based protocol. Read More

    Disulfide bond mapping of Pfs25, a recombinant malaria transmission blocking vaccine candidate.
    Anal Biochem 2017 Nov 21;542:20-23. Epub 2017 Nov 21.
    PATH Malaria Vaccine Initiative (MVI), 455 Massachusetts Avenue NW, Suite 1000, Washington, DC 20001-2621, USA. Electronic address:
    A liquid chromatography tandem-mass spectrometry method was developed to map the eleven disulfide bonds in Pfs25, a malaria transmission-blocking vaccine candidate. The compact and complex nature of Pfs25 has led to difficulties in prior peptide mapping efforts. Here, we report confirmation of proper disulfide pairing of a recombinant Pfs25, by optimizing denaturation and digestion with trypsin/Lys-C. Read More

    Development of a low-cost paper-based ELISA method for rapid Escherichia coli O157:H7 detection.
    Anal Biochem 2017 Nov 20;542:58-62. Epub 2017 Nov 20.
    Department of Hygienic Inspection, School of Public Health, Jilin University, Changchun 130021, Jilin, China. Electronic address:
    Escherichia coli O157: H7 (E. coli O157: H7) has become one of the most dangerous foodborne pathogenic bacteria around the world. Currently, because of the tedious, high-cost and stringent laboratory conditions required, the conventional E. Read More

    Mitochondrial transfer between cells: Methodological constraints in cell culture and animal models.
    Anal Biochem 2017 Nov 21. Epub 2017 Nov 21.
    School of Biological Sciences, Victoria University, PO Box 600, Wellington 6140, New Zealand.
    Interest in the recently discovered phenomenon of mitochondrial transfer between mammalian cells has gained momentum since it was first described in cell culture systems more than a decade ago. Mitochondria-targeting fluorescent dyes have been repurposed and are now widely used in these studies and in acute disease models, sometimes without due consideration of their limitations, while vectors containing mitochondrially-imported fluorescent proteins have complemented the use of mitochondria-targeting dyes. Genetic approaches that use mitochondrial DNA polymorphisms have also been used in some in vitro studies and in tumor models and are particularly useful where mtDNA is damaged or deleted. Read More

    Fullerene-PAMAM(G5) composite modified impedimetric biosensor to detect Fetuin-A in real blood samples.
    Anal Biochem 2017 Nov 20;542:11-15. Epub 2017 Nov 20.
    Ege University, Faculty of Medicine, Medical Biochemistry Department, Bornova, İzmir, Turkey.
    The aim of this study is to develop a nanomaterial-dendrimer composite modified biosensor to detect Fetuin-A (HFA) in real blood samples. For this purpose, we designed an Electrochemical Impedance Spectroscopy (EIS) based anti-Fetuin-A (Anti-HFA) modified biosensor system and tested in real blood samples. Chronoimpedance was also employed. Read More

    A multianalytical approach to investigate the effect of nanofiltration on plasma-derived factor IX clinical lots.
    Anal Biochem 2017 Nov 20;542:1-10. Epub 2017 Nov 20.
    Dipartimento di Scienze e Tecnologie Ambientali, Biologiche e Farmaceutiche, DISTABIF, Università della Campania Luigi Vanvitelli, Via Vivaldi, 43, 81100, Caserta, Italy. Electronic address:
    Plasma-derived proteins are a subset of relevant biotherapeutics also known as "well-characterized biologicals". They are enriched from plasma through several steps of physical and biochemical methodologies, reaching the regulatory accepted standards of safety, levels of impurities, activity and lot-to-lot consistency. Final products accepted for commercialization are submitted to tight analytical, functional and safety controls by a number of different approaches that fulfill the requirements of sensitivity and reliability. Read More

    Functionalization of paramagnetic nanoparticles for protein immobilization and purification.
    Anal Biochem 2018 Jan 20;540-541:45-51. Epub 2017 Nov 20.
    Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP CEP 14049-901, Brazil. Electronic address:
    A paramagnetic nanocomposite coated with chitosan and N-(5-Amino-1-carboxy-pentyl) iminodiacetic acid (NTA) that is suitable for protein immobilization applications has been prepared and characterized. The nanoparticle core was synthesized by controlled aggregation of Fe3O4 under alkaline conditions, and Transmission Electron Microscopy revealed a size distribution of 10-50 nm. The nanoparticle core was coated with chitosan and derivatized with glutaraldehyde and NTA, as confirmed by Fourier Transform Infrared Spectroscopy. Read More

    Circulating tumoral DNA: Preanalytical validation and quality control in a diagnostic laboratory.
    Anal Biochem 2017 Nov 11;542:34-39. Epub 2017 Nov 11.
    Molecular and Genomic Diagnostic Laboratory, Service of Medical Genetics, Department of Genetic Medicine, Laboratory Medicine and Pathology, University Hospitals of Geneva, CH-1211 Geneva, Switzerland. Electronic address:
    We present the results of our technical validation process in establishing the analysis of circulating tumor DNA (ctDNA) as a diagnostic tool. Like most cells in our body, tumor cells shed DNA in the blood flow. Analysis of ctDNA mutational content can provide invaluable information on the genetic makeup of a tumor, and assist oncologists in deciding on therapy, or in following residual disease. Read More

    Detection of SNPs of T2DM susceptibility genes by a ligase detection reaction-fluorescent nanosphere technique.
    Anal Biochem 2018 Jan 9;540-541:38-44. Epub 2017 Nov 9.
    Department of Endocrinology, The Second Hospital of Jilin University, Changchun 130000, China.
    Objective: To establish a high throughput, low cost, and simple nanotechnology-based method for the detection of single nucleotide polymorphism (SNP) loci in type 2 diabetes mellitus (T2DM).

    Methods: Multiplex ligase detection reaction (LDR) amplification was performed using fluorescently labeled magnetic nanosphere-bound upstream LDR probes and downstream probes labeled with a unique fluorescent group for each SNP locus. The amplified LDR products were separated by magnetic nanospheres and then scanned by fluorescence spectroscopy. Read More

    Antibody drug quantitation in coexistence with anti-drug antibodies on nSMOL bioanalysis.
    Anal Biochem 2018 Jan 9;540-541:30-37. Epub 2017 Nov 9.
    Leading Technology of Bioanalysis and Protein Chemistry, SHIMADZU Corporation, Kuwabara-cho, Nishino-kyo, Nakagyo-ku, Kyoto 604-8511, Japan. Electronic address:
    Therapeutic monoclonal antibodies (mAbs) are developed for treatment of diverse cancers and autoimmune diseases. For expansion of mAbs approval against unapproved diseases and pharmaceutical development, pharmacokinetics study is very important. Bioanalysis provides one of the most essential index against pharmacokinetics information. Read More

    A recombinant fusion protein-based, fluorescent protease assay for high throughput-compatible substrate screening.
    Anal Biochem 2018 Jan 6;540-541:52-63. Epub 2017 Nov 6.
    Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary. Electronic address:
    In connection with the intensive investigation of proteases, several methods have been developed for analysis of the substrate specificity. Due to the great number of proteases and the expected target molecules to be analyzed, time- and cost-efficient high-throughput screening (HTS) methods are preferred. Here we describe the development and application of a separation-based HTS-compatible fluorescent protease assay, which is based on the use of recombinant fusion proteins as substrates of proteases. Read More

    Label-free electrochemical immunoassay for neuron specific enolase based on 3D macroporous reduced graphene oxide/polyaniline film.
    Anal Biochem 2018 Jan 4;540-541:1-8. Epub 2017 Nov 4.
    College of Chemistry and Molecular Engineering, Nanjing Tech University, Nanjing 211816, PR China. Electronic address:
    The content of neuron specific enolase (NSE) in serum is considered to be an essential indicator of small cell lung cancer (SCLC). Here, a novel label-free electrochemical immunoassay for the detection of NSE based on the three dimensionally macroporous reduced graphene oxide/polyaniline (3DM rGO/PANI) film has been proposed. The 3DM rGO/PANI film was constructed by electrochemical co-deposition of GO and aniline into the interspaces of a sacrificial silica opal template modified Au slice. Read More

    Development of a quantitative immuno-polymerase chain reaction assay to detect and quantify low levels of human thyroid stimulating hormone.
    Anal Biochem 2017 Dec 27;539:134-143. Epub 2017 Oct 27.
    Instituto de Salud y Ambiente del Litoral (ISAL, CONICET-UNL), Ciudad Universitaria, Paraje El Pozo s/n, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, CP3000 Santa Fe, Argentina. Electronic address:
    In the present study, we developed both a conventional enzyme-linked immunosorbent assay (ELISA) and a highly sensitive immuno-polymerase chain reaction (IPCR) assay specific for detection of human thyroid stimulating hormone (hTSH). Several anti-hTSH monoclonal antibodies (MAbs) were generated using hybridoma technology. Two pairs of MAbs (B-4 and B-9) were rationally selected and the optimal assay conditions of sandwich ELISAs were established. Read More

    An ELISA method to estimate the mono ADP-ribosyltransferase activities: e.g in pertussis toxin and vaccines.
    Anal Biochem 2018 Jan 9;540-541:15-19. Epub 2017 Nov 9.
    TDI, National Institute for Biological Standards and Control (NIBSC), Blanche Lane, South Mimms, Potters Bar, Hertfordshire, EN6 3QG, UK.
    ADP-ribosyltransferase activities have been observed in many prokaryotic and eukaryotic species and viruses and are involved in many cellular processes, including cell signalling, DNA repair, gene regulation and apoptosis. In a number of bacterial toxins, mono ADP-ribosyltransferase is the main cause of host cell cytotoxicity. Several approaches have been used to analyse this biological system from measuring its enzyme products to its functions. Read More

    Detection of Pseudomonas aeruginosa biomarkers from thermally injured mice in situ using imaging mass spectrometry.
    Anal Biochem 2017 Dec 28;539:144-148. Epub 2017 Oct 28.
    Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT 59717, USA. Electronic address:
    Monitoring patients with burn wounds for infection is standard practice because failure to rapidly and specifically identify a pathogen can result in poor clinical outcomes, including death. Therefore, a method that facilitates detection and identification of pathogens in situ within minutes of biopsy would be a significant benefit to clinicians. Mass spectrometry is rapidly becoming a standard tool in clinical settings, capable of identifying specific pathogens from complex samples. Read More

    Facile detection of microRNA based on phosphorescence resonance energy transfer and duplex-specific nuclease-assisted signal amplification.
    Anal Biochem 2017 Dec 28;539:127-133. Epub 2017 Oct 28.
    Shanxi Normal University, Linfen, Shanxi 041000, China. Electronic address:
    MicroRNAs (miRNAs) play an important role in many biological processes, and its level in plasma and other biological fluids is closely related to many diseases. In this work, a selective room-temperature phosphorescence (RTP) detection method for miRNA was developed based on a duplex-specific nuclease (DSN) -assisted signal amplification strategy and phosphorescence resonance energy transfer (PRET) between poly-diallyldimethylammonium chloride-modified quantum dots (QDs@PDDA) and 6-carboxy-X-rhodamine-modified miRNA sequences complementary oligonucleotide (ROX-ssDNA). The positively charged QDs@PDDA could adsorb negatively charged ROX-ssDNA by electrostatic interaction, whereas the RTP signal of QDs@PDDA could be efficiently quenched by ROX-ssDNA via PRET. Read More

    Glycan profile of CHO derived IgM purified by highly efficient single step affinity chromatography.
    Anal Biochem 2017 Dec 4;539:162-166. Epub 2017 Nov 4.
    Department of Biotechnology, VIBT, University of Natural Resources and Life Sciences, Vienna, Muthgasse 18, 1190 Vienna, Austria. Electronic address:
    Immunoglobulin M (IgM) antibodies are reckoned as promising tools for therapy and diagnostic approaches. Nevertheless, the commercial success of IgMs is hampered due to bottlenecks in recombinant production and downstream processing. IgMs are large, complex and highly glycosylated proteins that are only stable in a limited range of conditions. Read More

    Quantification of sulphur amino acids by ultra-high performance liquid chromatography in aquatic invertebrates.
    Anal Biochem 2017 Dec 26;539:158-161. Epub 2017 Oct 26.
    Department of Biology & School of Geography and Earth Sciences, McMaster University, 1280 Main Street W., Hamilton, ON, L8S 4K1, Canada.
    We examined the performance of an ultra-high performance liquid chromatography method to quantify protein-bound sulphur amino acids in zooplankton. Both cysteic acid and methionine sulfone were linear from 5 to 250 pmol (r2 = 0.99), with a method detection limit of 13 pmol and 9 pmol, respectively. Read More

    Fluorometric determination of d-lactate in biological fluids.
    Anal Biochem 2017 Dec 3;539:152-157. Epub 2017 Nov 3.
    Dept. of Animal Science, Aarhus University, Blichers Allé 20, Postboks 50, DK-8830, Tjele, Denmark. Electronic address:
    Objective: D-lactic acid in the mammalian body is mainly of microbiological origin and is often located somewhere along the digestive tract. Surgical, extensive re-sectioning of the small bowel may be one of the risk factors for altered balance in the microbiological environment. Higher levels in the body may lead to D-lactate acidosis and neurotoxicity; consequently, the possibility of diagnosis of this condition is important. Read More

    Chelatable trace zinc causes low, irreproducible KDAC8 activity.
    Anal Biochem 2018 Jan 31;540-541:9-14. Epub 2017 Oct 31.
    Department of Chemistry, Xavier University of Louisiana, 1 Drexel Dr., New Orleans, LA 70125-1098, USA. Electronic address:
    Acetylation is an important regulatory mechanism in cells, and emphasis is being placed on identifying substrates and small molecule modulators of this post-translational modification. However, the reported in vitro activity of the lysine deacetylase KDAC8 is inconsistent across experimental setups, even with the same substrate, complicating progress in the field. We detected trace levels of zinc, a known inhibitor of KDAC8 when present in excess, even in high-quality buffer reagents, at concentrations that are sufficient to significantly inhibit the enzyme under common reaction conditions. Read More

    A novel voltammetric approach for real-time electrochemical detection of targeted nucleic acid sequences using LAMP.
    Anal Biochem 2017 Dec 24;539:113-117. Epub 2017 Oct 24.
    Toshiba Corporation, 1, Komukai-Toshiba-cho, Saiwai-ku, Kawasaki 212-8582, Japan.
    We have developed a novel voltammetric DNA chip for real-time electrochemical detection of targeted nucleic acid sequences using loop-mediated isothermal amplification (LAMP) and ruthenium hexaamine (RuHex) as the intercalative redox compound. A GspSSD DNA polymerase was used for LAMP owing to its tolerance of the intercalative redox compound. The electrochemical reaction of 1 mM RuHex in the LAMP solution was measured continuously by linear sweep voltammetry at 65 °C using an electrochemical DNA chip. Read More

    Design of titanium nitride- and wolfram carbide-doped RGO/GC electrodes for determination of gallic acid.
    Anal Biochem 2017 Dec 24;539:104-112. Epub 2017 Oct 24.
    The Vinča Institute of Nuclear Sciences, University of Belgrade, POB 522, 11001 Belgrade, Serbia.
    In the present paper, the electrochemical behavior and the properties of two modified glassy carbon (GC) electrodes used for quantification of gallic acid in sweet wines were compared. A comparative study was conducted between titanium nitride- or wolfram carbide-doped reduced graphene oxide, labeled as TNrGO and WCrGO, respectively, modified GC electrodes, which are promising composite nanomaterials for electroanalytical applications. For the first time, WCrGO was synthesized and its electroanalytical properties compared with those of TNrGO. Read More

    Helix structure of the double-stranded DNA for aptameric biosensing and imaging of cytochrome c.
    Anal Biochem 2018 Jan 23;540-541:20-29. Epub 2017 Oct 23.
    Department of Chemistry, Khorramabad Branch, Islamic Azad University, Khorramabad, Iran. Electronic address:
    Here, a method is introduced for construction the aptameric biosensor for biosensing detection of cytochrome C (CYC) based on chain-shape structure of aptasensor by using highly dispersed silver nanoparticles (AgNPs) on acid-oxidized carbon nanotube (CNTs) substrate. The animated capture probe (ssDNA1) and CYC-aptamer (ssDNA2) was immobilized on AgNPs/CNTs surface by covalent amide bonds formed by the carboxyl groups on the nanotubes and the amino groups on the oligonucleotides and hybridization, respectively. In this protocol, the nucleic acids at both ends of the ssDNA1 were sequenced to be complementary (tailor-made ssDNA1). Read More

    In situ surface protein conjugation of small molecules for SPR immunoassays.
    Anal Biochem 2017 Dec 21;539:149-151. Epub 2017 Oct 21.
    The New Zealand Institute for Plant & Food Research Limited, 120 Mt Albert Road, Sandringham, Auckland 1025, New Zealand. Electronic address:
    Solution based protein conjugation of small molecules involves multiple steps of chemical syntheses, bio-conjugation and purifications which are labor intensive and time-consuming. Since many small molecules have limited water solubility, conjugation to a protein is also a relatively low efficiency process in aqueous solutions. In this study, a model in situ protein conjugation of small molecules was achieved onto SPR surfaces, using progesterone and ovalbumin as a model small-molecule-protein system. Read More

    Quantifying variant differences in DNA melting curves: Effects of length, melting rate, and curve overlay.
    Anal Biochem 2017 Dec 21;539:90-95. Epub 2017 Oct 21.
    Department of Pathology, University of Utah School of Medicine, 383 Colorow Drive, Salt Lake City, UT 84108, USA. Electronic address:
    High resolution DNA melting of PCR products is a simple technique for sequence variant detection and analysis. However, sensitivity and specificity vary and depend on many factors that continue to be defined. We introduce the area between normalized melting curves as a metric to quantify genotype discrimination. Read More

    Electrochemical quantification of some water soluble vitamins in commercial multi-vitamin using poly-amino acid caped by graphene quantum dots nanocomposite as dual signal amplification elements.
    Anal Biochem 2017 Dec 19;539:70-80. Epub 2017 Oct 19.
    Department of Nano Technology, Nano Technology Research Center, Faculty of Chemistry, Urmia University, Urmia 57154, Iran; Pharmaceutical Analysis Research Center, Tabriz University of Medical Sciences, Tabriz 51664, Iran.
    Rapid analyses of some water soluble vitamins (Vitamin B2, B9, and C) in commercial multi vitamins could be routinely performed in analytical laboratories. This study reports on the electropolymerization of a low toxic and biocompatible polymer "poly aspartic acid-graphene quantum dots" as a novel strategy for surface modification of glassy carbon electrode and preparation a new interface for measurement of selected vitamins in commercial multi vitamins. Electrochemical deposition, as a well-controlled synthesis procedure, has been used for subsequently layer-by-layer preparation of graphene quantum dots nanostructures on a poly aspartic acid using cyclic voltammetry techniques in the regime of -1. Read More

    Development of amide-based fluorescent probes for selective measurement of carboxylesterase 1 activity in tissue extracts.
    Anal Biochem 2017 Dec 18;539:81-89. Epub 2017 Oct 18.
    Department of Entomology and Nematology, UC Davis Comprehensive Cancer Center, University of California, Davis, Davis, CA 95616, USA. Electronic address:
    Carboxylesterases are well known for their role in the metabolism of xenobiotics. However, recent studies have also implicated carboxylesterases in regulating a number of physiological processes including metabolic homeostasis and macrophage development, underlying the need to quantify them individually. Unfortunately, current methods for selectively measuring the catalytic activity of individual carboxylesterases are not sufficiently sensitive to support many biological studies. Read More

    Using two-site binding models to analyze microscale thermophoresis data.
    Anal Biochem 2018 Jan 18;540-541:64-75. Epub 2017 Oct 18.
    Department of Biophysics, The University of Texas Southwestern Medical Center, Dallas, TX, USA; Department of Microbiology, The University of Texas Southwestern Medical Center, Dallas, TX, USA. Electronic address:
    The emergence of microscale thermophoresis (MST) as a technique for determining the dissociation constants for bimolecular interactions has enabled these quantities to be measured in systems that were previously difficult or impracticable. However, most models for analyses of these data featured the assumption of a simple 1:1 binding interaction. The only model widely used for multiple binding sites was the Hill equation. Read More

    Determination of mitochondrial metabolic phenotype through investigation of the intrinsic inhibition of succinate dehydrogenase.
    Anal Biochem 2017 Oct 14. Epub 2017 Oct 14.
    Department of Surgery, Drexel University College of Medicine, Philadelphia, PA, USA.
    Many diseases are accompanied by systemic or organ metabolic abnormalities. Therefore, investigation of the roles of mitochondrial dysfunction in the pathogenesis of major diseases requires a methodology that reflects the characteristics of mitochondrial metabolism particular for the organ of origin. We provide evidence that for brain and heart mitochondria the intrinsic inhibition of succinate dehydrogenase (SDH) is a key mechanism for attenuation of mitochondrial respiration and energy production in response to the organ's energy needs. Read More

    Microvalve controlled multi-functional microfluidic chip for divisional cell co-culture.
    Anal Biochem 2017 Dec 12;539:48-53. Epub 2017 Oct 12.
    Beijing Key Laboratory of Bioseparation and Bioanalysis, Beijing Institute of Technology, Beijing 100081, China. Electronic address:
    Pneumatic micro-valve controlled microfluidic chip provides precise fluidic control for cell manipulation. In this paper, a multi-functional microfluidic chip was designed for three separate experiments: 1. Different cell lines were dispensed and cultured; 2. Read More

    Immunoaffinity capture coupled with capillary electrophoresis - mass spectrometry to study therapeutic protein stability in vivo.
    Anal Biochem 2017 Dec 10;539:118-126. Epub 2017 Oct 10.
    Pharmacokinetics and Drug Metabolism, Amgen Inc., 1120 Veterans Boulevard, South San Francisco, CA, USA.
    Protein engineering is at an all-time high in biopharmaceutics. As a result, absorption, distribution, metabolism and excretion (ADME) of proteins has become more important to understand in the context of engineering strategies to optimize therapeutic properties of potential lead constructs. Immunoaffinity capture coupled with a newly developed capillary electrophoresis - mass spectrometry (CE-MS) system was used to characterize intact protein mass analysis of a wild type Fc-FGF21 construct and a sequence re-engineered Fc-FGF21 construct from an in vivo study. Read More

    A multiplex RNA quantification method to determine the absolute amounts of mRNA without reverse transcription.
    Anal Biochem 2017 Dec 10;539:96-103. Epub 2017 Oct 10.
    Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo, 153-8902, Japan. Electronic address:
    We have developed a highly sensitive microarray-based method that determines the absolute amounts of mRNA in a total RNA sample in a multiplex manner without reverse transcription. This direct mRNA measurement promotes high-throughput testing and reduces bias in transcriptome analyses. Furthermore, quantification of the absolute amount of mRNA allows transcriptome analysis without common controls or additional, complicated normalization. Read More

    Spontaneous luminescence color change in the firefly luciferase assay system.
    Anal Biochem 2017 Dec 13;539:54-59. Epub 2017 Oct 13.
    Department of Applied Chemistry, National Chiao Tung University, 1001 University Rd, Hsinchu, 300, Taiwan; Department of Chemistry, National Tsing Hua University, 101, Section 2, Kuang-Fu Rd., Hsinchu, 30013, Taiwan. Electronic address:
    The temporal effects of luciferase reaction luminescence have only been discussed in the context of light intensity (flash vs. glow). However, alterations in the color of the light emitted over the course of the luciferase reaction have not been reported. Read More

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