Displaying publications 1 - 20 of 366 in total

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  1. Cheng Y, Lai OM, Tan CP, Panpipat W, Cheong LZ, Shen C
    ACS Appl Mater Interfaces, 2021 Jan 27;13(3):4146-4155.
    PMID: 33440928 DOI: 10.1021/acsami.0c17134
    Immobilization can be used to improve the stability of lipases and enhances lipase recovery and reusability, which increases its commercial value and industrial applications. Nevertheless, immobilization frequently causes conformational changes of the lipases, which decrease lipase catalytic activity. in the present work, we synthesized UIO-66 and grafted UIO-66 crystals with proline for immobilization of Candida rugosa lipase (CRL). As indicated by steady-state fluorescence microscopy, grafting of proline onto UIO-66 crystals induced beneficial conformational change in CRL. CRL immobilized on UIO-66/Pro (CRL@UIO-66/Pro) demonstrated higher enzyme activity and better recyclability than that immobilized on UIO-66 (CRL@UIO-66) in both hydrolysis (CRL@UIO-66/Pro: 0.34 U; CRL@UIO-66: 0.15 U) and transesterification (CRL@UIO-66/Pro: 0.93 U; CRL@UIO-66: 0.25 U) reactions. The higher values of kcat and kcat/Km of CRL@UIO-66/Pro also showed that it had better catalytic efficiency as compared to CRL@UIO-66. It is also worth noting that CRL@UIO-66/Pro (0.93 U) demonstrated a much higher transesterification activity as compared to free CRL (0.11 U), indicating that UIO-66/Pro has increased the solvent stability of CRL. Both CRL@UIO-66 and CRL@UIO-66/Pro were also used for the fabrication of biosensors for nitrofen with a wide linear range (0-100 μM), lower limit of detection, and good recovery rate.
    Matched MeSH terms: Biosensing Techniques/methods
  2. Ravikumar A, Panneerselvam P, Morad N
    ACS Appl Mater Interfaces, 2018 Jun 20;10(24):20550-20558.
    PMID: 29792319 DOI: 10.1021/acsami.8b05041
    In this paper, we propose a metal-polydopamine (MPDA) framework with a specific molecular probe which appears to be the most promising approach to a strong fluorescence quencher. The MPDA framework quenching ability toward various organic fluorophore such as aminoethylcoumarin acetate, 6-carboxyfluorescein (FAM), carboxyteramethylrhodamine, and Cy5 are used to establish a fluorescent biosensor that can selectively recognize Hg2+ and Ag+ ions. The fluorescent quenching efficiency was sufficient to achieve more than 96%. The MPDA framework also exhibits different affinities with ssDNA and dsDNA. In addition, the FAM-labeled ssDNA was adsorbed onto the MPDA framework, based on their interaction with the complex formed between MPDA frameworks/ssDNA taken as a sensing platform. By taking advantage of this sensor, highly sensitive and selective determination of Hg2+ and Ag+ ions is achieved through exonuclease III signal amplification activity. The detection limits of Hg2+ and Ag+ achieved to be 1.3 and 34 pM, respectively, were compared to co-existing metal ions and graphene oxide-based sensors. Furthermore, the potential applications of this study establish the highly sensitive fluorescence detection targets in environmental and biological fields.
    Matched MeSH terms: Biosensing Techniques
  3. Zhang J, Lakshmipriya T, Gopinath SCB
    ACS Omega, 2020 Oct 13;5(40):25899-25905.
    PMID: 33073115 DOI: 10.1021/acsomega.0c03260
    The primary reasons for myocardial infarction (MI) are pericarditis, arrhythmia, and heart failure, causing predominant deaths worldwide. Patients need a potential diagnostic system and treatment before cardiomyocyte damage. Cardiac biomarkers are released from myocytes immediately after a heart attack. Troponin is an efficient biomarker released from dead cells within a few hours. Aptamers are artificial antibodies used effectively in the biosensor field for biomarker detection. Along with aptamers, the application of nanomaterials is also expected to enhance the detection limits of biosensors. In this investigation, selected aptamers against cardiac troponin I (cTnI) were conjugated with gold nanoparticles (GNPs) to diagnose MI and compared with an aptamer-only control group on an interdigitated electrode surface. Based on electroanalysis, cTnI was detected at concentrations as low as 1 fM, and the detection limit improved to 100 aM when the aptamer was conjugated with GNP. In addition, aptamer-GNP conjugates increased the current level at the tested concentrations of cTnI. Control experiments with noncomplementary aptamers and relevant proteins did not result in notable changes in the current, demonstrating the selective detection of cTnI.
    Matched MeSH terms: Biosensing Techniques
  4. Tsiamis A, Diaz Sanchez F, Hartikainen N, Chung M, Mitra S, Lim YC, et al.
    ACS Omega, 2021 Apr 27;6(16):10568-10577.
    PMID: 34056211 DOI: 10.1021/acsomega.0c05823
    This paper presents a scalable method of developing ultrasensitive electrochemical biosensors. This is achieved by maximizing sensor conductivity through graphene wrapping of carbonized electrospun nanofibers. The effectiveness of the graphene wrap was determined visually by scanning electron microscopy and chemically by Fourier transform infrared spectroscopy, Raman spectroscopy, and X-ray diffraction. The sensing performance of different electrode samples was electrochemically characterized using cyclic voltammetry and electrochemical impedance spectroscopy, with the graphene-wrapped carbonized nanofiber electrode showing significantly improved performance. The graphene-wrapped carbonized nanofibers exhibited a relative conductivity of ∼14 times and an electroactive surface area of ∼2 times greater compared to the bare screen-printed carbon electrode despite experiencing inhibitive effects from the carbon glue used to bind the samples to the electrode. The results indicate potential for a highly conductive, inert sensing platform.
    Matched MeSH terms: Biosensing Techniques
  5. Ashley J, Shukor Y, D'Aurelio R, Trinh L, Rodgers TL, Temblay J, et al.
    ACS Sens, 2018 02 23;3(2):418-424.
    PMID: 29333852 DOI: 10.1021/acssensors.7b00850
    Food recalls due to undeclared allergens or contamination are costly to the food manufacturing industry worldwide. As the industry strives for better manufacturing efficiencies over a diverse range of food products, there is a need for the development of new analytical techniques to improve monitoring of the presence of unintended food allergens during the food manufacturing process. In particular, the monitoring of wash samples from cleaning in place systems (CIP), used in the cleaning of food processing equipment, would allow for the effective removal of allergen containing ingredients in between food batches. Casein proteins constitute the biggest group of proteins in milk and hence are the most common milk protein allergen in food ingredients. As such, these proteins could present an ideal analyte for cleaning validation. In this work, molecularly imprinted polymer nanoparticles (nanoMIPs) with high affinity toward bovine α-casein were synthesized using a solid-phase imprinting method. The nanoMIPs were then characterized and incorporated into label free surface plasmon resonance (SPR) based sensor. The nanoMIPs demonstrated good binding affinity and selectivity toward α-casein (KD ∼ 10 × 10-9 M). This simple affinity sensor demonstrated the quantitative detection of α-casein achieving a detection limit of 127 ± 97.6 ng mL-1 (0.127 ppm) which is far superior to existing commercially available ELISA kits. Recoveries from spiked CIP wastewater samples were within the acceptable range (87-120%). The reported sensor could allow food manufacturers to adequately monitor and manage food allergen risk in food processing environments while ensuring that the food produced is safe for the consumer.
    Matched MeSH terms: Biosensing Techniques/instrumentation; Biosensing Techniques/methods*
  6. Issa, R., Hamdan, N.A., Raj, A.S.S., Noh, M.F.M.
    ASM Science Journal, 2011;5(1):36-42.
    MyJurnal
    Researchers have developed and modified DNA biosensor techniques to provide a fast, simple and sensitive method for detection of human diseases, bacterial food contamination, forensic and environmental research. This study describes the physical characterization of screen-printed carbon electrodes using the scanning electron microscope.
    Matched MeSH terms: Biosensing Techniques
  7. Mohd Zain Z, Ab Ghani S, O'Neill RD
    Amino Acids, 2012 Nov;43(5):1887-94.
    PMID: 22865247 DOI: 10.1007/s00726-012-1365-0
    This paper discusses the application of a reagentless, selective microbiosensor as a useful alternative tool for monitoring D-serine in neural samples. The main components of the 125-μm-diameter disk biosensor were D-amino acid oxidase for D-serine sensitivity (linear region slope, 61 ± 7 μA cm(-2) mM(-1); limit of detection, 20 nM), and poly-phenylenediamine for rejection of electroactive interference. The response time of the biosensor was of the order of 1 s, ideal for 'real-time' monitoring, and detection of systemically administered D-serine in brain extracellular fluid is demonstrated. Exploitation of this probe might resolve queries involving regulation of D-serine in excitotoxicity, and modulation of N-methyl-D-aspartate receptor function by D-serine and glycine in the central nervous system.
    Matched MeSH terms: Biosensing Techniques*
  8. Nurul Najian AB, Engku Nur Syafirah EA, Ismail N, Mohamed M, Yean CY
    Anal Chim Acta, 2016 Jan 15;903:142-8.
    PMID: 26709307 DOI: 10.1016/j.aca.2015.11.015
    In recent years extensive numbers of molecular diagnostic methods have been developed to meet the need of point-of-care devices. Efforts have been made towards producing rapid, simple and inexpensive DNA tests, especially in the diagnostics field. We report on the development of a label-based lateral flow dipstick for the rapid and simple detection of multiplex loop-mediated isothermal amplification (m-LAMP) amplicons. A label-based m-LAMP lateral flow dipstick assay was developed for the simultaneous detection of target DNA template and a LAMP internal control. This biosensor operates through a label based system, in which probe-hybridization and the additional incubation step are eliminated. We demonstrated this m-LAMP assay by detecting pathogenic Leptospira, which causes the re-emerging disease Leptospirosis. The lateral flow dipstick was developed to detect of three targets, the LAMP target amplicon, the LAMP internal control amplicon and a chromatography control. Three lines appeared on the dipstick, indicating positive results for all representative pathogenic Leptospira species, whereas two lines appeared, indicating negative results, for other bacterial species. The specificity of this biosensor assay was 100% when it was tested with 13 representative pathogenic Leptospira species, 2 intermediate Leptospira species, 1 non-pathogenic Leptospira species and 28 other bacteria species. This study found that this DNA biosensor was able to detect DNA at concentrations as low as 3.95 × 10(-1) genomic equivalent ml(-1). An integrated m-LAMP and label-based lateral flow dipstick was successfully developed, promising simple and rapid visual detection in clinical diagnostics and serving as a point-of-care device.
    Matched MeSH terms: Biosensing Techniques
  9. Acquah C, Danquah MK, Yon JL, Sidhu A, Ongkudon CM
    Anal Chim Acta, 2015 Aug 12;888:10-8.
    PMID: 26320953 DOI: 10.1016/j.aca.2015.05.050
    The discovery of Systematic Evolution of Ligands by Exponential Enrichment (SELEX) assay has led to the generation of aptamers from libraries of nucleic acids. Concomitantly, aptamer-target recognition and its potential biomedical applications have become a major research endeavour. Aptamers possess unique properties that make them superior biological receptors to antibodies with a plethora of target molecules. Some specific areas of opportunities explored for aptamer-target interactions include biochemical analysis, cell signalling and targeting, biomolecular purification processes, pathogen detection and, clinical diagnosis and therapy. Most of these potential applications rely on the effective immobilisation of aptamers on support systems to probe target species. Hence, recent research focus is geared towards immobilising aptamers as oligosorbents for biodetection and bioscreening. This article seeks to review advances in immobilised aptameric binding with associated successful milestones and respective limitations. A proposal for high throughput bioscreening using continuous polymeric adsorbents is also presented.
    Matched MeSH terms: Biosensing Techniques/instrumentation; Biosensing Techniques/methods*
  10. Ngamdee T, Yin LS, Vongpunsawad S, Poovorawan Y, Surareungchai W, Lertanantawong B
    Anal Chim Acta, 2020 Oct 16;1134:10-17.
    PMID: 33059855 DOI: 10.1016/j.aca.2020.08.018
    DNA strand displacement is an attractive, enzyme-free target hybridization strategy for nano-biosensing. The target DNA induces a strand displacement reaction by replacing the pre-hybridized strand that is labeled with gold nanoparticles (AuNPs). Thus, the amount of displaced-AuNP-labeled strand is proportional to the amount of target DNA in the sample. The use of a magnetogenosensing technique to isolate the target DNA allows for a simple, one-pot detection approach, which minimizes possible carry-over contamination and pipetting errors. We sought a proof-of-concept for this technology in its ability to detect DNA-equivalent of hepatitis E virus (HEV), which causes acute viral hepatitis for which rapid and simple diagnostic methods remain limited. Signal detection was done via visual observation, spectrophotometry, and electrochemistry. The sensor demonstrated good sensitivity with detection limits of 10 pM (visual), 10 pM (spectrophotometry) and 1 fM (electrochemical). This sensor also exhibited high specificity for real target amplicons and could discriminate between perfect and mismatched sequences. Lyophilized biosensor reagents stored at 4 °C, 25 °C, and outdoor ambient temperature, were stable for up to 90, 50, and 40 days, respectively. The integration of magnetic separation and target DNA-induced strand displacement reaction in a dry reagent form makes the sensing platform easy-to-use and suitable for field settings.
    Matched MeSH terms: Biosensing Techniques*
  11. Wong XY, Quesada-González D, Manickam S, Muthoosamy K
    Anal Chim Acta, 2021 Aug 29;1175:338745.
    PMID: 34330444 DOI: 10.1016/j.aca.2021.338745
    Metal ions homeostasis plays an important role in biological processes. The ability to detect the concentration of metal ions in biological fluids is often challenged by the obvious interference or competitive binding nature of other alkaline metals ions. Common analytical techniques employed for metal ions detection are electrochemical, fluorescence and colorimetric methods. However, most reported metal ions sensors are complicated, time-consuming and involve costly procedures with limited effectiveness. Herein, a nanobiosensor for detecting sodium and potassium ions using folic acid-functionalised reduced graphene oxide-modified RNase A gold nanoclusters (FA-rGO-RNase A/AuNCs) based on fluorescence "turn-off/turn-on" is presented. Firstly, a facile and optimised protocol for the fabrication of RNase A/AuNCs is developed. The activity of RNase A protein after the formation of RNase A/AuNCs is studied. RNase A/AuNCs is then loaded onto FA-rGO, in which FA-rGO is used as a potential carrier and fluorescence quencher for RNase A/AuNCs. Finally, a fluorescence "turn-on" sensing strategy is developed using the as-synthesised FA-rGO-RNase A/AuNCs to detect sodium and potassium ions. The developed nanobiosensor revealed an excellent sensing performance and meets the sensitivity required to detect both sodium and potassium ions. To the best of our knowledge, this is the first work done on determining the RNase A protein activity in RNase A/AuNCs and exploring the potential application of RNase A/AuNCs as a metal ion sensor. This work serves as a proof-of-concept for combining the potential of drug delivery, active targeting and therapy on cancer cells, as well as biosensing of metal ions into a single platform.
    Matched MeSH terms: Biosensing Techniques*
  12. Fathil MF, Md Arshad MK, Ruslinda AR, Nuzaihan M N M, Gopinath SC, Adzhri R, et al.
    Anal Chim Acta, 2016 Sep 07;935:30-43.
    PMID: 27543013 DOI: 10.1016/j.aca.2016.06.012
    A real-time ability to interpret the interaction between targeted biomolecules and the surface of semiconductors (metal transducers) into readable electrical signals, without biomolecular modification involving fluorescence dyes, redox enzymes, and radioactive labels, created by label-free biosensors has been extensively researched. Field-effect transistor (FET)- and capacitor-based biosensors are among the diverse electrical charge biosensing architectures that have drawn much attention for having charge transduction; thus, enabling the early and rapid diagnosis of the appropriate cardiac biomarkers at lower concentrations. These semiconducting material-based transducers are very suitable to be integrated with portable electronic devices for future online collection, transmission, reception, analysis, and reporting. This overview elucidates and clarifies two major electrical label-free systems (FET- and capacitor-based biosensors) with cardiac troponin (cTn) biomarker-mediated charge transduction for acute myocardial infarction (AMI) diagnosis. Advances in these systems are highlighted by their progression in bridging the laboratory and industry; the foremost technologies have made the transition from benchtop to bedside and beyond.
    Matched MeSH terms: Biosensing Techniques*
  13. Foo PC, Chan YY, Mohamed M, Wong WK, Nurul Najian AB, Lim BH
    Anal Chim Acta, 2017 May 08;966:71-80.
    PMID: 28372729 DOI: 10.1016/j.aca.2017.02.019
    This study highlighted the development of a four target nitrocellulose-based nucleic acid lateral flow immunoassay biosensor in a dry-reagent strip format for interpretation of double-labelled double-stranded amplicons from thermostabilised triplex loop-mediated isothermal amplification assay. The DNA biosensor contained two test lines which captured biotin and texas red labelled amplicons; a LAMP internal amplification control line that captured digoxigenin labelled amplicon; and a chromatography control line that validated the functionality of the conjugated gold nanoparticles and membrane. The red lines on detection pad were generated when the gold nanoparticles conjugated antibody bound to the fluorescein labelled amplicons, and the capture agents bound to their specific hapten on the other 5' end of the double-stranded amplicon. The applicability of this DNA biosensor was demonstrated using amoebiasis-causing Entamoeba histolytica simultaneously with the non-pathogenic but morphologically identical Entamoeba dispar and Entamoeba moshkovskii. The biosensor detection limit was 10 E. histolytica trophozoites, and revealed 100% specificity when it was evaluated against 3 medically important Entamoeba species and 75 other pathogenic microorganisms. Heat stability test showed that the biosensor was stable for at least 181 days at ambient temperature. This ready-to-use and cold-chain-free biosensor facilitated the post-LAMP analysis based on visualisation of lines on strip instead of observation of amplicon patterns in agarose gel.
    Matched MeSH terms: Biosensing Techniques*
  14. Weihs F, Peh A, Dacres H
    Anal Chim Acta, 2020 Mar 15;1102:99-108.
    PMID: 32044001 DOI: 10.1016/j.aca.2019.12.044
    Proteases are key signalling molecules for many physiological processes and their dysregulation is implicated in the progression of a range of diseases. Sensitive methods to measure protease activities in complex biological samples are critical for rapid disease diagnoses. The proteolytic activity of plasmin reflects the fibrinolysis state of blood and its deregulation can indicate pathologies such as bleeding events. While Bioluminescence Resonance Energy Transfer (BRET) is a powerful and sensitive method for the detection of protease activity, the commonly applied blue-shifted BRET2 system, consisting of the Renilla luciferase Rluc2 and the large-stokes shift fluorescent protein GFP2, suffers from light absorption and light scattering in human plasma samples. To address this challenge, we developed a red-shifted BRET-based plasmin sensor by substituting BRET2 with the BRET6 system. BRET6 is composed of the red-shifted RLuc8.6 luciferase linked to the red light emitting fluorescent protein TurboFP635. The BRET6 biosensor exhibited 3-fold less light absorption in plasma samples compared to the BRET2 sensor leading to an up to a 5-fold increase in sensitivity for plasmin detection in plasma. The limits of detection for plasmin were determined to be 11.90 nM in 7.5% (v/v) plasma with a 10 min assay which enables biologically relevant plasmin activities of thrombolytic therapies to be detected. While a colorigenic plasmin activity assay achieved a similar detection limit of 10.91 nM in 7.5% (v/v) human plasma, it required a 2 h incubation period. The BRET6 sensor described here is faster and more specific than the colorigenic assay as it did not respond to unspiked human plasma samples.
    Matched MeSH terms: Biosensing Techniques/methods
  15. Chen TY, Mani V, Huang ST, Chang PC, Huang CH, Huang NM
    Anal Chim Acta, 2017 Oct 16;990:78-83.
    PMID: 29029745 DOI: 10.1016/j.aca.2017.08.051
    An electrochemical latent redox probe, SAF 5 was designed, synthesized and characterized. A rapid and sensitive solution-based assay was demonstrated for salicylate hydroxylase (SHL). In presence of NADH at aerobic conditions, SHL catalyzed the decarboxylative hydroxylation of SAF and released a redox reporter amino ferrocene (AF 6). The release of AF 6 was monitored at interference free potential region (-50 mV vs. Ag|AgCl) using differential pulse voltammetry as signal read-out. The current signal generated by this process is highly specific, and insensitive to other biological interfering compounds. Next, the SAF incorporated SHL assay was extended to fabricate immobilization-free biosensors for rapid sensing of salicylic acid (SA) and β-hydroxybutyrate (β-HB) in whole blood. The described method rapidly detects SA in a linear range of 35-560 μM with detection limit of 5.0 μM. For β-HB determination, the linear range was 10-600 μM and detection limit was 2.0 μM. Besides, the assay protocols are simple, fast, reliable, selective, sensitive and advantageous over existing methods. The whole blood assay did not required cumbersome steps such as, enzyme immobilization, pre-treatments and holds great practical potential in clinical diagnosis.
    Matched MeSH terms: Biosensing Techniques*
  16. Yew CT, Azari P, Choi JR, Li F, Pingguan-Murphy B
    Anal Chim Acta, 2018 Jun 07;1009:81-88.
    PMID: 29422135 DOI: 10.1016/j.aca.2018.01.016
    Point-of-care biosensors are important tools developed to aid medical diagnosis and testing, food safety and environmental monitoring. Paper-based biosensors, especially nucleic acid-based lateral flow assays (LFA), are affordable, simple to produce and easy to use in remote settings. However, the sensitivity of such assays to infectious diseases has always been a restrictive challenge. Here, we have successfully electrospun polycaprolactone (PCL) on nitrocellulose (NC) membrane to form a hydrophobic coating to reduce the flow rate and increase the interaction rate between the targets and gold nanoparticles-detecting probes conjugates, resulting in the binding of more complexes to the capture probes. With this approach, the sensitivity of the PCL electrospin-coated test strip has been increased by approximately ten-fold as compared to the unmodified test strip. As a proof of concept, this approach holds great potential for sensitive detection of targets at point-of-care testing.
    Matched MeSH terms: Biosensing Techniques*
  17. Lee ST, Beaumont D, Su XD, Muthoosamy K, New SY
    Anal Chim Acta, 2018 Jun 20;1010:62-68.
    PMID: 29447672 DOI: 10.1016/j.aca.2018.01.012
    Single strand DNA (ssDNA) chimeras consisting of a silver nanoclusters-nucleating sequence (NC) and an aptamer are widely employed to synthesize functional silver nanoclusters (AgNCs) for sensing purpose. Despite its simplicity, this chimeric-templated AgNCs often leads to undesirable turn-off effect, which may suffer from false positive signals caused by interference. In our effort to elucidate how the relative position of NC and aptamer affects the fluorescence behavior and sensing performance, we systematically formulated these NC and aptamer regions at different position in a DNA chimera. Using adenosine aptamer as a model, we tested the adenosine-induced optical response of each design. We also investigated the effect of linker region connecting NC and aptamer, as well as different NC sequence on the sensing performance. We concluded that locating NC sequence at 5'-end exhibited the best response, with immediate fluorescence enhancement observed over a wide linear range (1-2500 μM). Our experimental findings help to explain the emission behavior and sensing performance of chimeric conjugates of AgNCs, providing an important means to formulate a better aptasensor.
    Matched MeSH terms: Biosensing Techniques
  18. Abdullah J, Ahmad M, Heng LY, Karuppiah N, Sidek H
    Anal Bioanal Chem, 2006 Nov;386(5):1285-92.
    PMID: 17031625
    The stacked-film immobilization of 3-methyl-2-benzothiazolinone hydrazone (MBTH) in hybrid nafion/sol-gel silicate film and horseradish peroxidase (HRP) in chitosan, performed in order to allow the determination of phenolic compounds, was investigated via an optical method. The stacked films were deposited onto a microscope glass slide by a spin-coating technique. The quinone or free radical product formed by the enzymatic reactions of phenolic compounds interacts with MBTH to form azo-dye products, which can be measured spectrophotometrically at a wavelength of 500 nm. The color intensity of the product was found to increase in proportion to the phenolic concentration after 5 min of exposure. The response of the biosensor was linear over concentration ranges of 0.025-0.500, 0.010-0.070 and 0.050-0.300 mM for guaiacol, resorcinol and o-cresol, respectively, and gave detection limits of 0.010, 0.005 and 0.012 mM. The sensor exhibited good sensitivity and stability for at least two months.
    Matched MeSH terms: Biosensing Techniques/methods
  19. Azri FA, Selamat J, Sukor R, Yusof NA, Raston NHA, Eissa S, et al.
    Anal Bioanal Chem, 2021 Jun;413(15):3861-3872.
    PMID: 34021369 DOI: 10.1007/s00216-021-03336-1
    Aptamers are short single-stranded oligonucleotides (either DNA or RNA) that can fold into well-defined three-dimensional (3D) spatial structures which enable them to capture their specific target by complementary shape interactions. Aptamers are selected from large random libraries through the SELEX process and only a small fraction of the sequence is involved in direct docking with the target. In this paper, we describe the possible truncation variants of zearalenone (ZEA) aptamer which might be an effective binding region for the target. The originally selected zearalenone (ZEA) aptamer was 80-mer in length and shown to bind the target with a high affinity (Kd = 41 ± 5 nM). Herein, computational docking simulation was performed with 15 truncated variants to determine the predicted binding energy and responsible binding site of the aptamer-analyte complex. The results revealed that 5 truncated variants had binding energy lower than - 7.0 kcal/mol. Circular dichroism analysis was performed on the shortlisted aptamer and the conformational change of aptamers was observed with the presence of an analyte. Aptamer Z3IN (29-mer) was chosen as the most enhanced affinity for its target with a dissociation constant of 11.77 ± 1.44 nM. The aptamer was further applied in the electrochemical aptasensor of ZEA based on an indirect competitive format. The results demonstrated that the truncated aptamer leads to an enhancement of the sensitivity of the biosensor.
    Matched MeSH terms: Biosensing Techniques/methods
  20. Muniandy S, Dinshaw IJ, Teh SJ, Lai CW, Ibrahim F, Thong KL, et al.
    Anal Bioanal Chem, 2017 Nov;409(29):6893-6905.
    PMID: 29030671 DOI: 10.1007/s00216-017-0654-6
    Reduced graphene oxide (rGO) has emerged as a promising nanomaterial for reliable detection of pathogenic bacteria due to its exceptional properties such as ultrahigh electron transfer ability, large surface to volume ratio, biocompatibility, and its unique interactions with DNA bases of the aptamer. In this study, rGO-azophloxine (AP) nanocomposite aptasensor was developed for a sensitive, rapid, and robust detection of foodborne pathogens. Besides providing an excellent conductive and soluble rGO nanocomposite, the AP dye also acts as an electroactive indicator for redox reactions. The interaction of the label-free single-stranded deoxyribonucleic acid (ssDNA) aptamer with the test organism, Salmonella enterica serovar Typhimurium (S. Typhimurium), was monitored by differential pulse voltammetry analysis, and this aptasensor showed high sensitivity and selectivity for whole-cell bacteria detection. Under optimum conditions, this aptasensor exhibited a linear range of detection from 108 to 101 cfu mL-1 with good linearity (R 2 = 0.98) and a detection limit of 101 cfu mL-1. Furthermore, the developed aptasensor was evaluated with non-Salmonella bacteria and artificially spiked chicken food sample with S. Typhimurium. The results demonstrated that the rGO-AP aptasensor possesses high potential to be adapted for the effective and rapid detection of a specific foodborne pathogen by an electrochemical approach. Graphical abstract Fabrication of graphene-based nanocomposite aptasensor for detection of foodborne pathogen.
    Matched MeSH terms: Biosensing Techniques/instrumentation*; Biosensing Techniques/methods*
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