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  1. Formulation of DNA chimera templates: Effects on emission behavior of silver nanoclusters and sensing
    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: Aptamers, Nucleotide/chemistry
  2. SELEX Modifications and Bioanalytical Techniques for Aptamer-Target Binding Characterization
    Tan SY, Acquah C, Sidhu A, Ongkudon CM, Yon LS, Danquah MK
    Crit Rev Anal Chem, 2016 Nov;46(6):521-37.
    PMID: 26980177 DOI: 10.1080/10408347.2016.1157014
    The quest to improve the detection of biomolecules and cells in health and life sciences has led to the discovery and characterization of various affinity bioprobes. Libraries of synthetic oligonucleotides (ssDNA/ssRNA) with randomized sequences are employed during Systematic Evolution of Ligands by Exponential Enrichment (SELEX) to select highly specific affinity probes called aptamers. With much focus on the generation of aptamers for a variety of target molecules, conventional SELEX protocols have been modified to develop new and improved SELEX protocols yielding highly specific and stable aptamers. Various techniques have been used to analyze the binding interactions between aptamers and their cognate molecules with associated merits and limitations. This article comprehensively reviews research advancements in the generation of aptamers, analyses physicochemical conditions affecting their binding characteristics to cellular and biomolecular targets, and discusses various field applications of aptameric binding. Biophysical techniques employed in the characterization of the molecular and binding features of aptamers to their cognate targets are also discussed.
    Matched MeSH terms: Aptamers, Nucleotide/chemistry*
  3. Aptamers: an emerging class of bioaffinity ligands in bioactive peptide applications
    Acquah C, Agyei D, Obeng EM, Pan S, Tan KX, Danquah MK
    Crit Rev Food Sci Nutr, 2020;60(7):1195-1206.
    PMID: 30714390 DOI: 10.1080/10408398.2018.1564234
    The food and health applications of bioactive peptides have grown remarkably in the past few decades. Current elucidations have shown that bioactive peptides have unique structural arrangement of amino acids, conferring distinct functionalities, and molecular affinity characteristics. However, whereas interest in the biological potency of bioactive peptides has grown, cost-effective techniques for monitoring the structural changes in these peptides and how these changes affect the biological properties have not grown at the same rate. Due to the high binding affinity of aptamers for other biomolecules, they have a huge potential for use in tracking the structural, conformational, and compositional changes in bioactive peptides. This review provides an overview of bioactive peptides and their essential structure-activity relationship. The review further highlights on the types and methods of synthesis of aptamers before the discussion of the prospects, merits, and challenges in the use of aptamers for bioaffinity interactions with bioactive peptides.
    Matched MeSH terms: Aptamers, Nucleotide/chemistry*
  4. Assays for aptamer-based platforms
    Citartan M, Gopinath SC, Tominaga J, Tan SC, Tang TH
    Biosens Bioelectron, 2012 Apr 15;34(1):1-11.
    PMID: 22326894 DOI: 10.1016/j.bios.2012.01.002
    Aptamers are single stranded DNA or RNA oligonucleotides that have high affinity and specificity towards a wide range of target molecules. Aptamers have low molecular weight, amenable to chemical modifications and exhibit stability undeterred by repetitive denaturation and renaturation. Owing to these indispensable advantages, aptamers have been implemented as molecular recognition element as alternative to antibodies in various assays for diagnostics. By amalgamating with a number of methods that can provide information on the aptamer-target complex formation, aptamers have become the elemental tool for numerous biosensor developments. In this review, administration of aptamers in applications involving assays of fluorescence, electrochemistry, nano-label and nano-constructs are discussed. Although detection strategies are different for various aptamer-based assays, the core of the design strategies is similar towards reporting the presence of specific target binding to the corresponding aptamers. It is prognosticated that aptamers will find even broader applications with the development of new methods of transducing aptamer target binding.
    Matched MeSH terms: Aptamers, Nucleotide/chemistry*
  5. Recent developments of aptasensors expedient for point-of-care (POC) diagnostics
    Citartan M, Tang TH
    Talanta, 2019 Jul 01;199:556-566.
    PMID: 30952298 DOI: 10.1016/j.talanta.2019.02.066
    Aptamers are nucleic acid-based molecular recognition elements that are specific and have high binding affinity against their respective targets. On account of their target recognition capacity, aptamers are widely utilized in a number of applications including diagnostics. This review aims to highlight the recent developments of aptasensors expedient for point-of-care (POC) diagnostics. Significant focus is given on the primary assay formats of aptamers such as fluorescence, electrochemical, surface plasmon resonance (SPR) and colorimetric assays. A potpourri of platforms such as paper-based device, lateral flow assay, portable electrodes, portable SPR and smart phones expedient for point-of-care (POC) diagnostics are discussed. Emphasis is also given on the technicalities and assay configurations associated with the sensors.
    Matched MeSH terms: Aptamers, Nucleotide/chemistry*
  6. Aptamer-17β-estradiol-antibody sandwich ELISA for determination of gynecological endocrine function
    Huang Y, Zhang L, Li Z, Gopinath SCB, Chen Y, Xiao Y
    Biotechnol Appl Biochem, 2021 Aug;68(4):881-888.
    PMID: 33245588 DOI: 10.1002/bab.2008
    17β-Estradiol-E2 (17β-E2) is a steroid hormone that plays a major role in the reproductive endocrine system and is involved in various processes, such as pregnancy, fertility, and menopause. In this study, the performance of an enzyme-linked immunosorbent assay (ELISA) for 17β-E2 quantification was enhanced by using a gold nanoparticle (GNP)-conjugated aptamer. An anti-17β-E2-aptamer-GNP antibody was immobilized on an amine-modified ELISA surface. Then, 17β-E2 was allowed to interact with and be sandwiched by antibodies. Aptamer-GNP conjugation was confirmed by colorimetric assays via the naked eye and UV-visible light spectroscopy. The detection limit based on a signal-to-noise ratio (S/N) of 3 was estimated to be 1.5 nM (400 pg/mL), and the linear range was 1.5-50 nM. Control experiments (without 17β-E2/with a complementary aptamer sequence/with a nonimmune antibody) confirmed the specific detection of 17β-E2. Moreover, 17β-E2 spiking of human serum did not interrupt the interaction between 17β-E2 and its antibody and aptamer. Thus, the developed ELISA can be used as an alternate assay for quantification of 17β-E2 and assessment of endocrine-related gynecological problems.
    Matched MeSH terms: Aptamers, Nucleotide/chemistry*
  7. In vitro selection and characterization of single stranded DNA aptamers for luteolin: A possible recognition tool
    Tuma Sabah J, Zulkifli RM, Shahir S, Ahmed F, Abdul Kadir MR, Zakaria Z
    Anal Biochem, 2018 05 15;549:72-79.
    PMID: 29524380 DOI: 10.1016/j.ab.2018.03.004
    Distinctive bioactivities possessed by luteolin (3', 4', 5, 7-tetrahydroxy-flavone) are advantageous for sundry practical applications. This paper reports the in vitro selection and characterization of single stranded-DNA (ssDNA) aptamers, specific for luteolin (LUT). 76-mer library containing 1015 randomized ssDNA were screened via systematic evolution of ligands by exponential enrichment (SELEX). The recovered ssDNA pool from the 8th round was amplified with unlabeled primers and cloned into PSTBlue-1 vector prior to sequencing. 22 of LUT-binding aptamer variants were further classified into one of the seven groups based on their N40 random sequence regions, wherein one representative from each group was characterized. The dissociation constant of aptamers designated as LUT#28, LUT#20 and LUT#3 was discerned to be 107, 214 and 109 nM, respectively with high binding affinity towards LUT. Prediction analysis of the secondary structure suggested discrete features with typical loop and stem motifs. Furthermore, LUT#3 displayed higher specificity with insignificant binding toward kaempferol and quercetin despite its structural and functional similarity compared to LUT#28 and LUT#20. Further LUT#3 can detect free luteolin within 0.2-1 mM in solution. It was suggested that LUT#3 aptamer were the most suitable for LUT recognition tool at laboratory scale based on the condition tested.
    Matched MeSH terms: Aptamers, Nucleotide/chemistry
  8. Aptamers as a replacement for antibodies in enzyme-linked immunosorbent assay
    Toh SY, Citartan M, Gopinath SC, Tang TH
    Biosens Bioelectron, 2015 Feb 15;64:392-403.
    PMID: 25278480 DOI: 10.1016/j.bios.2014.09.026
    The application of antibodies in enzyme-linked immunosorbent assay (ELISA) is the basis of this diagnostic technique which is designed to detect a potpourri of complex target molecules such as cell surface antigens, allergens, and food contaminants. However, development of the systematic evolution of Ligands by Exponential Enrichment (SELEX) method, which can generate a nucleic acid-based probe (aptamer) that possess numerous advantages compared to antibodies, offers the possibility of using aptamers as an alternative molecular recognition element in ELISA. Compared to antibodies, aptamers are smaller in size, can be easily modified, are cheaper to produce, and can be generated against a wide array of target molecules. The application of aptamers in ELISA gives rise to an ELISA-derived assay called enzyme-linked apta-sorbent assay (ELASA). As with the ELISA method, ELASA can be used in several different configurations, including direct, indirect, and sandwich assays. This review provides an overview of the strategies involved in aptamer-based ELASA.
    Matched MeSH terms: Aptamers, Nucleotide/chemistry*
  9. A review on immobilised aptamers for high throughput biomolecular detection and screening
    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: Aptamers, Nucleotide/chemistry*
  10. High-performance interactive analysis of split aptamer and HIV-1 Tat on multiwall carbon nanotube-modified field-effect transistor
    Fatin MF, Rahim Ruslinda A, Gopinath SCB, Arshad MKM
    Int J Biol Macromol, 2019 Mar 15;125:414-422.
    PMID: 30529550 DOI: 10.1016/j.ijbiomac.2018.12.066
    Interaction between split RNA aptamer and the clinically important target, HIV-1 Tat was investigated on a biosensing surface transduced by functionally choreographed multiwall carbon nanotubes (MWCNTs). Acid oxidation was performed to functionalize MWCNTs with carboxyl functional groups. X-ray photoelectron spectroscopy analysis had profound ~2.91% increment in overall oxygen group and ~1% increment was noticed with a specific carboxyl content owing to CO and OCO bonding. The interaction between split RNA aptamer and HIV-1 Tat protein was quantified by electrical measurements with the current signal (Ids) over a gate voltage (Vgs). Initially, 34.4 mV gate voltage shift was observed by the immobilization of aptamer on MWCNT. With aptamer and HIV-1 Tat interaction, the current flow was decreased with the concomitant gate voltage shift of 23.5 mV. The attainment of sensitivity with split aptamer and HIV-1 Tat interaction on the fabricated device was 600 pM. To ensure the genuine interaction of aptamer with HIV-1 Tat, other HIV-1 proteins, Nef and p24 were interacted with aptamer and they displayed the negligible interferences with gate voltage shift of 3.5 mV and 5.7 mV, which shows 4 and 2.5 folds lesser than HIV-1 Tat interaction, respectively.
    Matched MeSH terms: Aptamers, Nucleotide/chemistry*
  11. Graphene oxide-gold nanoparticle-aptamer complexed probe for detecting amyloid beta oligomer by ELISA-based immunoassay
    Zhao J, Chang W, Liu L, Xing X, Zhang C, Meng H, et al.
    J Immunol Methods, 2021 02;489:112942.
    PMID: 33333060 DOI: 10.1016/j.jim.2020.112942
    Highly sensitive and easy detection method for Alzheimer's disease (AD) with a suitable biomarker is mandatory for preventing the factors resulting from AD. This research reports a modified ELISA with graphene for the detection of AD biomarker amyloid beta (Aβ) oligomer. Gold nanoparticle (AuNP) conjugated aptamer was used as the capture probe and attached on ELISA-graphene oxide surface through the amine linker. Antibody was used as the detection molecule to reach the maximum detection of Aβ oligomer. Suitable level of APTMS (2%), size of AuNP (30 nm) and aptamer concentration (2 μM) were optimized. This sandwich pattern of aptamer-Aβ oligomer-antibody helps to reach the detection at 50 pM on the optimized ELISA surface and the control experiments in the absence of Aβ oligomer or anti-Aβ oligomer antibody did not show the significant optical detection at 492 nm, indicting the specific detection. Further, Aβ oligomer spiked artificial cerebrospinal fluid did not interfere the detection of Aβ oligomer, confirming the selective detection. This new and modified ELISA surface helps to reach the lower detection of Aβ oligomer and diagnose AD.
    Matched MeSH terms: Aptamers, Nucleotide/chemistry*
  12. Biomolecular assembly on interdigitated electrode nanosensor for selective detection of insulin-like growth factor-1
    Gu Y, Liu L, Guo J, Xiao S, Fang F, Yu X, et al.
    Artif Cells Nanomed Biotechnol, 2021 Dec;49(1):30-37.
    PMID: 33467925 DOI: 10.1080/21691401.2020.1865992
    This research is focussed to quantify IGF1 by electroanalytical analysis on InterDigitated electrode surface and characterized by the microscopic observations. For the detection, antibody and aptamer were used to analyze the level of IGF1. The sandwich pattern (aptamer-IGF1-antibody) was designed on the chemically modified IDE surface and reached the limit of detection to 10 fM with 100 folds enhancement in the sensitivity. Different control experiments (absence of IGF1, binding with IGF2 and with non-complementary aptamer) were failed to show the current changes, discriminated the specific detection. A good detection strategy is to complement the currently following imaging systems for AAA.
    Matched MeSH terms: Aptamers, Nucleotide/chemistry*
  13. Determination of minimal sequence for zearalenone aptamer by computational docking and application on an indirect competitive electrochemical aptasensor
    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: Aptamers, Nucleotide/chemistry
  14. Diagnosing human blood clotting deficiency
    Ong CC, Gopinath SCB, Rebecca LWX, Perumal V, Lakshmipriya T, Saheed MSM
    Int J Biol Macromol, 2018 Sep;116:765-773.
    PMID: 29775720 DOI: 10.1016/j.ijbiomac.2018.05.084
    There are different clotting factors present in blood, carries the clotting cascade and excessive bleeding may cause a deficiency in the clotting Diagnosis of this deficiency in clotting drastically reduces the potential fatality. For enabling a sensor to detect the clotting factors, suitable probes such as antibody and aptamer have been used to capture these targets on the sensing surface. Two major clotting factors were widely studied for the diagnosis of clotting deficiency, which includes factor IX and thrombin. In addition, factor IX is considered as the substitute for heparin and the prothrombotic associated with the increased thrombin generation are taking into account their prevalence. The biosensors, surface plasmon resonance, evanescent-field-coupled waveguide-mode sensor, metal-enhanced PicoGreen fluorescence and electrochemical aptasensor were well-documented and improvements have been made for high-performance sensing. We overviewed detecting factor IX and thrombin using these biosensors, for the potential application in medical diagnosis.
    Matched MeSH terms: Aptamers, Nucleotide/chemistry*
  15. Molecularly imprinted polymer amalgamation on narrow-gapped Archimedean-spiral interdigitated electrodes: resistance to electrolyte fouling in acidic medium
    Krishnan H, Gopinath SCB, Md Arshad MK, Zulhaimi HI, Ramanathan S
    Mikrochim Acta, 2021 03 31;188(4):144.
    PMID: 33791872 DOI: 10.1007/s00604-021-04794-1
    A conventional photolithography technique was used to fabricate three types of Archimedean-spiral interdigitated electrodes (AIDEs) containing concentric interlocking electrodes with different electrode and gap sizes, i.e., 150 μm (D1), 100 μm (D2), and 50 μm (D3). The precision of the fabrication was validated by surface topography using scanning electron microscopy, high power microscopy, 3D-nano profilometry, and atomic force microscopy. These AIDEs were fabricated with a tolerance of ± 6 nm in dimensions. The insignificant current variation at the pico-ampere range for all bare AIDEs further proved the reproducibility of the device. The large gap sized AIDE (D1) is insensitive to acidic medium, whereas D2 and D3 are insensitive to alkali medium. D2 was the best with regard to its electrical characterization. Furthermore, uniformly synthesized molecularly imprinted polymer (MIP) nanoparticles prepared with human blood clotting factor IX and its aptamer were in the size range 140 to 160 nm, attached on the sensing surface and characterized. The average thickness of deposited MIP film was 1.7 μm. EDX data shows the prominent peaks for silicon and aluminum substrates as 61.79 and 22.52%, respectively. The MIP nanoparticles-deposited sensor surface was characterized by applying it in electrolyte solutions, and smooth curves with the current flow were observed at pH lower than 8 and discriminated against alkali media. This study provides a new MIP amalgamated AIDE with nano-gapped fingers enabling analysis of other biomaterials due to its operation in an ideal buffer range.
    Matched MeSH terms: Aptamers, Nucleotide/chemistry
  16. Fulltext 1,1'-Carbonyldiimidazole-copper nanoflower enhanced collapsible laser scribed graphene engraved microgap capacitive aptasensor for the detection of milk allergen
    Subramani IG, Perumal V, Gopinath SCB, Mohamed NM, Ovinis M, Sze LL
    Sci Rep, 2021 10 21;11(1):20825.
    PMID: 34675227 DOI: 10.1038/s41598-021-00057-4
    The bovine milk allergenic protein, 'β-lactoglobulin' is one of the leading causes of milk allergic reaction. In this research, a novel label-free non-faradaic capacitive aptasensor was designed to detect β-lactoglobulin using a Laser Scribed Graphene (LSG) electrode. The graphene was directly engraved into a microgapped (~ 95 µm) capacitor-electrode pattern on a flexible polyimide (PI) film via a simple one-step CO2 laser irradiation. The novel hybrid nanoflower (NF) was synthesized using 1,1'-carbonyldiimidazole (CDI) as the organic molecule and copper (Cu) as the inorganic molecule via one-pot biomineralization by tuning the reaction time and concentration. NF was fixed on the pre-modified PI film at the triangular junction of the LSG microgap specifically for bio-capturing β-lactoglobulin. The fine-tuned CDI-Cu NF revealed the flower-like structures was viewed through field emission scanning electron microscopy. Fourier-transform infrared spectroscopy showed the interactions with PI film, CDI-Cu NF, oligoaptamer and β-lactoglobulin. The non-faradaic sensing of milk allergen β-lactoglobulin corresponds to a higher loading of oligoaptamer on 3D-structured CDI-Cu NF, with a linear range detection from 1 ag/ml to 100 fg/ml and attomolar (1 ag/ml) detection limit (S/N = 3:1). This novel CDI-Cu NF/LSG microgap aptasensor has a great potential for the detection of milk allergen with high-specificity and sensitivity.
    Matched MeSH terms: Aptamers, Nucleotide/chemistry*
  17. Graphene-based label-free electrochemical aptasensor for rapid and sensitive detection of foodborne pathogen
    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: Aptamers, Nucleotide/chemistry
  18. Prospects in the use of aptamers for characterizing the structure and stability of bioactive proteins and peptides in food
    Agyei D, Acquah C, Tan KX, Hii HK, Rajendran SRCK, Udenigwe CC, et al.
    Anal Bioanal Chem, 2018 Jan;410(2):297-306.
    PMID: 28884330 DOI: 10.1007/s00216-017-0599-9
    Food-derived bioactive proteins and peptides have gained acceptance among researchers, food manufacturers and consumers as health-enhancing functional food components that also serve as natural alternatives for disease prevention and/or management. Bioactivity in food proteins and peptides is determined by their conformations and binding characteristics, which in turn depend on their primary and secondary structures. To maintain their bioactivities, the molecular integrity of bioactive peptides must remain intact, and this warrants the study of peptide form and structure, ideally with robust, highly specific and sensitive techniques. Short single-stranded nucleic acids (i.e. aptamers) are known to have high affinity for cognate targets such as proteins and peptides. Aptamers can be produced cost-effectively and chemically derivatized to increase their stability and shelf life. Their improved binding characteristics and minimal modification of the target molecular signature suggests their suitability for real-time detection of conformational changes in both proteins and peptides. This review discusses the developmental progress of systematic evolution of ligands by exponential enrichment (SELEX), an iterative technology for generating cost-effective aptamers with low dissociation constants (Kd) for monitoring the form and structure of bioactive proteins and peptides. The review also presents case studies of this technique in monitoring the structural stability of bioactive peptide formulations to encourage applications in functional foods. The challenges and potential of aptamers in this research field are also discussed. Graphical abstract Advancing bioactive proteins and peptide functionality via aptameric ligands.
    Matched MeSH terms: Aptamers, Nucleotide/chemistry*
  19. Aptamer-Mediated Polymeric Vehicles for Enhanced Cell-Targeted Drug Delivery
    Tan KX, Danquah MK, Sidhu A, Yon LS, Ongkudon CM
    Curr Drug Targets, 2018 02 08;19(3):248-258.
    PMID: 27321771 DOI: 10.2174/1389450117666160617120926
    BACKGROUND: The search for smart delivery systems for enhanced pre-clinical and clinical pharmaceutical delivery and cell targeting continues to be a major biomedical research endeavor owing to differences in the physicochemical characteristics and physiological effects of drug molecules, and this affects the delivery mechanisms to elicit maximum therapeutic effects. Targeted drug delivery is a smart evolution essential to address major challenges associated with conventional drug delivery systems. These challenges mostly result in poor pharmacokinetics due to the inability of the active pharmaceutical ingredients to specifically act on malignant cells thus, causing poor therapeutic index and toxicity to surrounding normal cells. Aptamers are oligonucleotides with engineered affinities to bind specifically to their cognate targets. Aptamers have gained significant interests as effective targeting elements for enhanced therapeutic delivery as they can be generated to specifically bind to wide range of targets including proteins, peptides, ions, cells and tissues. Notwithstanding, effective delivery of aptamers as therapeutic vehicles is challenged by cell membrane electrostatic repulsion, endonuclease degradation, low pH cleavage, and binding conformation stability.

    OBJECTIVE: The application of molecularly engineered biodegradable and biocompatible polymeric particles with tunable features such as surface area and chemistry, particulate size distribution and toxicity creates opportunities to develop smart aptamer-mediated delivery systems for controlled drug release.

    RESULTS: This article discusses opportunities for particulate aptamer-drug formulations to advance current drug delivery modalities by navigating active ingredients through cellular and biomolecular traffic to target sites for sustained and controlled release at effective therapeutic dosages while minimizing systemic cytotoxic effects.

    CONCLUSION: A proposal for a novel drug-polymer-aptamer-polymer (DPAP) design of aptamer-drug formulation with stage-wise delivery mechanism is presented to illustrate the potential efficacy of aptamer- polymer cargos for enhanced cell targeting and drug delivery.

    Matched MeSH terms: Aptamers, Nucleotide/chemistry
  20. Single-stranded DNA (ssDNA) production in DNA aptamer generation
    Marimuthu C, Tang TH, Tominaga J, Tan SC, Gopinath SC
    Analyst, 2012 Mar 21;137(6):1307-15.
    PMID: 22314701 DOI: 10.1039/c2an15905h
    The discovery that synthetic short chain nucleic acids are capable of selective binding to biological targets has made them to be widely used as molecular recognition elements. These nucleic acids, called aptamers, are comprised of two types, DNA and RNA aptamers, where the DNA aptamer is preferred over the latter due to its stability, making it widely used in a number of applications. However, the success of the DNA selection process through Systematic Evolution of Ligands by Exponential Enrichment (SELEX) experiments is very much dependent on its most critical step, which is the conversion of the dsDNA to ssDNA. There is a plethora of methods available in generating ssDNA from the corresponding dsDNA. These include asymmetric PCR, biotin-streptavidin separation, lambda exonuclease digestion and size separation on denaturing-urea PAGE. Herein, different methods of ssDNA generation following the PCR amplification step in SELEX are reviewed.
    Matched MeSH terms: Aptamers, Nucleotide/chemistry
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