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A High-Throughput Magnetic Nanoparticle-Based Semi-Automated Antibody Phage Display Biopanning

Ch'ng ACW, Ahmad A, Konthur Z, Lim TS

Methods Mol Biol, 2019;1904:377-400.
PMID: 30539481 DOI: 10.1007/978-1-4939-8958-4_18

Panning is a common process used for antibody selection from phage antibody libraries. There are several methods developed for a similar purpose, namely streptavidin mass spectrometry immunoassay (MSIA™) Disposable Automation Research Tips, magnetic beads, polystyrene immunotubes, and microtiter plate. The advantage of using a magnetic particle processor system is the ability to carry out phage display panning against multiple target antigens simultaneously in parallel. The system carries out the panning procedure using magnetic nanoparticles in microtiter plates. The entire incubation, wash, and elution process is then automated in this setup. The system also allows customization for the introduction of different panning stringencies. The nature of the biopanning process coupled with the limitation of the system means that minimal human intervention is required for the infection and phage packaging stage. However, the process still allows for rapid and reproducible antibody generation to be carried out.

Matched MeSH terms: High-Throughput Screening Assays*
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: High-Throughput Screening Assays/instrumentation; High-Throughput Screening Assays/methods
Fulltext Evaluation of extraction kits and RT-qPCR systems adapted to high-throughput platform for circulating miRNAs

Tan GW, Khoo AS, Tan LP

Sci Rep, 2015;5:9430.
PMID: 25800946 DOI: 10.1038/srep09430

MicroRNAs regulate gene expression at the post-transcriptional level. Differential expression of miRNAs can potentially be used as biomarkers for early diagnosis and prediction for outcomes. Failure in validation of miRNA profiles is often caused by variations in experimental parameters. In this study, the performance of five extraction kits and three RT-qPCR systems were evaluated using BioMark high-throughput platform and the effects of different experimental parameters on circulating miRNA levels were determined. Differences in the performance of extraction kits as well as varying accuracy, sensitivity and reproducibility in qPCR systems were observed. Normalisation of RT-qPCR data to spike-in controls can reduce extraction bias. However, the extent of correlation for different qPCR systems varies in different assays. At different time points, there was no significant fold change in eight of the plasma miRNAs that we evaluated. Higher level of miRNAs was detected in plasma as compared to serum of the same cohort. In summary, we demonstrated that high-throughput RT-qPCR with pre-amplification step had increased sensitivity and can be achieved with accuracy and high reproducibility through stringent experimental controls. The information provided here is useful for planning biomarker validation studies involving circulating miRNAs.

Matched MeSH terms: High-Throughput Screening Assays
Fulltext Cell-Based High-Throughput Screening Protocol for Discovering Antiviral Inhibitors Against SARS-COV-2 Main Protease (3CLpro)

Rothan HA, Teoh TC

Mol Biotechnol, 2021 Mar;63(3):240-248.
PMID: 33464543 DOI: 10.1007/s12033-021-00299-7

The global public health has been compromised since the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) emerged in late December 2019. There are no specific antiviral drugs available to combat SARS-CoV-2 infection. Besides the rapid dissemination of SARS-CoV-2, several variants have been identified with a potential epidemiologic and pathogenic variation. This fact has forced antiviral drug development strategies to stay innovative, including new drug discovery protocols, combining drugs, and establishing new drug classes. Thus, developing novel screening methods and direct-targeting viral enzymes could be an attractive strategy to combat SARS-CoV-2 infection. In this study, we designed, optimized, and validated a cell-based assay protocol for high-throughput screening (HTS) antiviral drug inhibitors against main viral protease (3CLpro). We applied the split-GFP complementation to develop GFP-split-3CLpro HTS system. The system consists of GFP-based reporters that become fluorescent upon cleavage by SARS-CoV-2 protease 3CLpro. We generated a stable GFP-split-3CLpro HTS system valid to screen large drug libraries for inhibitors to SARS-CoV-2 main protease in the bio-safety level 2 laboratory, providing real-time antiviral activity of the tested compounds. Using this assay, we identified a new class of viral protease inhibitors derived from quinazoline compounds that worth further in vitro and in vivo validation.

Matched MeSH terms: High-Throughput Screening Assays/methods*
High-Throughput RT-qPCR for the Analysis of Circulating MicroRNAs

Tan GW, Tan LP

Methods Mol Biol, 2017;1580:7-19.
PMID: 28439823 DOI: 10.1007/978-1-4939-6866-4_2

Reverse transcription followed by real-time or quantitative polymerase chain reaction (RT-qPCR) is the gold standard for validation of results from transcriptomic profiling studies such as microarray and RNA sequencing. The current need for most studies, especially biomarker studies, is to evaluate the expression levels or fold changes of many transcripts in a large number of samples. With conventional low to medium throughput qPCR platforms, many qPCR plates would have to be run and a significant amount of RNA input per sample will be required to complete the experiments. This is particularly challenging when the size of study material (small biopsy, laser capture microdissected cells, biofluid, etc.), time, and resources are limited. A sensitive and high-throughput qPCR platform is therefore optimal for the evaluation of many transcripts in a large number of samples because the time needed to complete the entire experiment is shortened and the usage of lab consumables as well as RNA input per sample are low. Here, the methods of high-throughput RT-qPCR for the analysis of circulating microRNAs are described. Two distinctive qPCR chemistries (probe-based and intercalating dye-based) can be applied using the methods described here.

Matched MeSH terms: High-Throughput Screening Assays/methods
Methods in Screening Antiviral Drugs Against Enterovirus 71

Bajaber NAOA, Ramanathan B

Methods Mol Biol, 2021;2296:167-184.
PMID: 33977447 DOI: 10.1007/978-1-0716-1358-0_9

Enteroviruses 71 (EV71) is a single-stranded, neurotrophic RNA virus responsible for the numerous outbreaks of hand, foot, and mouth disease (HFMD) in the Asia-Pacific regions. HFMD primarily affects children to cause range of infection, from mild symptoms to acute flaccid paralysis, and hemorrhage. Despite increased incidence of EV71 epidemics globally and research against EV71 becoming prioritized, no antiviral agent against EV71 has yet been licensed and approved worldwide. In this chapter, detailed EV71 antiviral screening techniques are described, including plaque assay which determines viral titers through the use of a semisolid overlay, carboxymethyl cellulose to allow even viral spread and infection across the host cellular monolayers as well as a crystal violet, a distinct counterstain to visualize circular regions of infectious zones-plaques. qRT-PCR is used to quantify the viral genomic RNA in the infected samples and MTS cell viability assay to quantify the cell viability after infection or toxicity of the compound on the cells. Furthermore, various antiviral inhibition assays including prophylactic, post infection, and virucidal assays are demonstrated for estimation of the antiviral activity of potential antiviral drugs against EV71. These methods can be effectively utilized in virology laboratories for effective high-throughput screening of antiviral molecules against EV71 that can assist in the future development of antiviral drugs.

Matched MeSH terms: High-Throughput Screening Assays/methods*
Fulltext Development of multicellular tumor spheroid (MCTS) culture from breast cancer cell and a high throughput screening method using the MTT assay

Ho WY, Yeap SK, Ho CL, Rahim RA, Alitheen NB

PLoS One, 2012;7(9):e44640.
PMID: 22970274 DOI: 10.1371/journal.pone.0044640

In comparison to monolayer cells, MCTS has been claimed as more suitable candidate for studying drug penetration due to the high resemblance to solid tumors. However, the cultivation of MCTS is cumbersome, time consuming, and most technique fail to generate spheroids with uniform sizes. Therefore, the application of spheroid cultures in high throughput screening has been rather limiting. Besides, the lack of a well established screening protocol method that is applicable to spheroid could also be attributed to this limitation. Here we report a simple way of cultivating homogenous MCTS cultures with compact and rigid structure from the MCF-7 cells. Besides, we had also made some modifications to the standard MTT assay to realize high throughput screening of these spheroids. Using the modified protocol, tamoxifen showed cytotoxicity effect towards MCTS cultures from MCF-7 with high consistency. The results correlated well with the cultures' response assessed by LDH release assay but the latter assay was not ideal for detecting a wide range of cytotoxicity due to high basal background reading. The MTT assay emerged as a better indicator to apoptosis event in comparison to the LDH release assay. Therefore, the method for spheroid generation and the modified MTT assay we reported here could be potentially applied to high throughput screening for response of spheroid cultures generated from MCF-7 as well as other cancer cell lines towards cytotoxic stimuli.

Matched MeSH terms: High-Throughput Screening Assays*
Fulltext High throughput molecular confirmation of β-thalassemia mutations using novel TaqMan probes

Kho SL, Chua KH, George E, Tan JA

Sensors (Basel), 2013;13(2):2506-14.
PMID: 23429513 DOI: 10.3390/s130202506

β-Thalassemia is a public health problem where 4.5% of Malaysians are β-thalassemia carriers. The genetic disorder is caused by defects in the β-globin gene complex which lead to reduced or complete absence of β-globin chain synthesis. Five TaqMan genotyping assays were designed and developed to detect the common β-thalassemia mutations in Malaysian Malays. The assays were evaluated with 219 "blinded" DNA samples and the results showed 100% sensitivity and specificity. The in-house designed TaqMan genotyping assays were found to be cost- and time-effective for characterization of β-thalassemia mutations in the Malaysian population.

Matched MeSH terms: High-Throughput Screening Assays/methods*
Whole-genome multiparametric screening to identify modulators of epithelial-to-mesenchymal transition

Said NA, Gould CM, Lackovic K, Simpson KJ, Williams ED

Assay Drug Dev Technol, 2014 Sep;12(7):385-94.
PMID: 25181411 DOI: 10.1089/adt.2014.593

Metastasis accounts for the poor prognosis of the majority of solid tumors. The phenotypic transition of nonmotile epithelial tumor cells to migratory and invasive "mesenchymal" cells (epithelial-to-mesenchymal transition [EMT]) enables the transit of cancer cells from the primary tumor to distant sites. There is no single marker of EMT; rather, multiple measures are required to define cell state. Thus, the multiparametric capability of high-content screening is ideally suited for the comprehensive analysis of EMT regulators. The aim of this study was to generate a platform to systematically identify functional modulators of tumor cell plasticity using the bladder cancer cell line TSU-Pr1-B1 as a model system. A platform enabling the quantification of key EMT characteristics, cell morphology and mesenchymal intermediate filament vimentin, was developed using the fluorescent whole-cell-tracking reagent CMFDA and a fluorescent promoter reporter construct, respectively. The functional effect of genome-wide modulation of protein-coding genes and miRNAs coupled with those of a collection of small-molecule kinase inhibitors on EMT was assessed using the Target Activation Bioapplication integrated in the Cellomics ArrayScan platform. Data from each of the three screens were integrated to identify a cohort of targets that were subsequently examined in a validation assay using siRNA duplexes. Identification of established regulators of EMT supports the utility of this screening approach and indicated capacity to identify novel regulators of this plasticity program. Pathway analysis coupled with interrogation of cancer-related expression profile databases and other EMT-related screens provided key evidence to prioritize further experimental investigation into the molecular regulators of EMT in cancer cells.

Matched MeSH terms: High-Throughput Screening Assays/methods*
Fulltext Dexterous robotic manipulation of alert adult Drosophila for high-content experimentation

Savall J, Ho ET, Huang C, Maxey JR, Schnitzer MJ

Nat. Methods, 2015 Jul;12(7):657-60.
PMID: 26005812 DOI: 10.1038/nmeth.3410

We present a robot that enables high-content studies of alert adult Drosophila by combining operations including gentle picking; translations and rotations; characterizations of fly phenotypes and behaviors; microdissection; or release. To illustrate, we assessed fly morphology, tracked odor-evoked locomotion, sorted flies by sex, and dissected the cuticle to image neural activity. The robot's tireless capacity for precise manipulations enables a scalable platform for screening flies' complex attributes and behavioral patterns.

Matched MeSH terms: High-Throughput Screening Assays*
A novel high throughput method based on the DPPH dry reagent array for determination of antioxidant activity

Musa KH, Abdullah A, Kuswandi B, Hidayat MA

Food Chem, 2013 Dec 15;141(4):4102-6.
PMID: 23993591 DOI: 10.1016/j.foodchem.2013.06.112

A stable chromogenic radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) is commonly used for the determination of antioxidant activity. In this paper, DPPH was dried into 96 well microplate to produce DPPH dry reagent array plate, based on which the highly sensitive and high throughput determination of antioxidant activities was achieved. The spectrophotometric characterization of the microplate containing dried or fresh DPPH free radicals was reported. The response of the DPPH dry reagent array towards different standard antioxidants was studied. The reaction for DPPH in fresh or dry reagent array with Trolox was reported and compared. The DPPH dry reagent array was used to study the antioxidant activity of banana, green tea, pink guava, and honeydew and the results were compared to the samples reacted with freshly prepared DPPH. The proposed method is comparable to the classical DPPH method, more convenient, simple to operate with minimal solvent required and excellent sensitivity.

Matched MeSH terms: High-Throughput Screening Assays/methods*
Current progress in production of flavonoids using systems and synthetic biology platforms

Ku Nurul Aqmar Ku Bahaudin, Ahmad Bazli Ramzi, Syarul Nataqain Baharum, Suriana Sabri, Adam Leow Thean Chor, Tewin Tencomnao

Sains Malaysiana, 2018;47:3077-3084.

Flavonoid is an industrially-important compound due to its high pharmaceutical and cosmeceutical values. However,
conventional methods in extracting and synthesizing flavonoids are costly, laborious and not sustainable due to small
amount of natural flavonoids, large amounts of chemicals and space used. Biotechnological production of flavonoids
represents a viable and sustainable route especially through the use of metabolic engineering strategies in microbial
production hosts. In this review, we will highlight recent strategies for the improving the production of flavonoids
using synthetic biology approaches in particular the innovative strategies of genetically-encoded biosensors for in
vivo metabolite analysis and high-throughput screening methods using fluorescence-activated cell sorting (FACS).
Implementation of transcription factor based-biosensor for microbial flavonoid production and integration of systems
and synthetic biology approaches for natural product development will also be discussed.

Matched MeSH terms: High-Throughput Screening Assays
Fulltext Elucidation of the compatible interaction between banana and Meloidogyne incognita via high-throughput proteome profiling

Al-Idrus A, Carpentier SC, Ahmad MT, Panis B, Mohamed Z

PLoS One, 2017;12(6):e0178438.
PMID: 28575037 DOI: 10.1371/journal.pone.0178438

With a diverse host range, Meloidogyne incognita (root-knot nematode) is listed as one of the most economically important obligate parasites of agriculture. This nematode species establishes permanent feeding sites in plant root systems soon after infestation. A compatible host-nematode interaction triggers a cascade of morphological and physiological process disruptions of the host, leading to pathogenesis. Such disruption is reflected by altered gene expression in affected cells, detectable using molecular approaches. We employed a high-throughput proteomics approach to elucidate the events involved in a compatible banana- M. incognita interaction. This study serves as the first crucial step in developing natural banana resistance for the purpose of biological-based nematode management programme. We successfully profiled 114 Grand naine root proteins involved in the interaction with M. incognita at the 30th- and 60th- day after inoculation (dai). The abundance of proteins involved in fundamental biological processes, cellular component organisation and stress responses were significantly altered in inoculated root samples. In addition, the abundance of proteins in pathways associated with defence and giant cell maintenance in plants such as phenylpropanoid biosynthesis, glycolysis and citrate cycle were also implicated by the infestation.

Matched MeSH terms: High-Throughput Screening Assays
Building a discovery partnership with Sarawak Biodiversity Centre: a gateway to access natural products from the rainforests

Yeo TC, Naming M, Manurung R

Comb Chem High Throughput Screen, 2014 Mar;17(3):192-200.
PMID: 24409959

The Sarawak Biodiversity Centre (SBC) is a state government agency which regulates research and promotes the sustainable use of biodiversity. It has a program on documentation of traditional knowledge (TK) and is well-equipped with facilities for natural product research. SBC maintains a Natural Product Library (NPL) consisting of local plant and microbial extracts for bioprospecting. The NPL is a core discovery platform for screening of bioactive compounds by researchers through a formal agreement with clear benefit sharing obligations. SBC aims to develop partnerships with leading institutions and the industries to explore the benefits of biodiversity.

Matched MeSH terms: High-Throughput Screening Assays
Rethinking zebrafish anxiety models in drug discovery and precision psychiatry

Abdul Basit NS, Yap MMC, Mah YK, Ogawa S

Expert Opin Drug Discov, 2026 Nov;21(11):1227-1241.
PMID: 42847163 DOI: 10.1080/17460441.2026.2743766

INTRODUCTION: Anxiety disorders are among the most prevalent psychiatric conditions worldwide, yet the development of novel anxiolytic therapies remains limited by poor translational success and the complexity of anxiety neurobiology. Zebrafish (Danio rerio) have emerged as a valuable vertebrate model bridging high-throughput in vitro screening and mammalian preclinical studies by combining whole-organism physiology with scalable phenotypic screening.
AREAS COVERED: This review evaluates advances in zebrafish models for anxiolytic drug discovery, focusing on features supporting their construct and predictive validity. Larval zebrafish enable high-throughput behavioral and imaging-based screening, whereas adults provide more complex behavioral phenotypes and translational validation. Genetically engineered models, automated behavioral tracking, functional imaging, and molecular profiling further support mechanism-based drug discovery. A targeted search of PubMed/MEDLINE, Web of Science, and Scopus identified current and foundational studies on zebrafish anxiety models, neurobiology, behavioral assays, and drug discovery.
EXPERT OPINION: Integration of behavioral phenotyping with genetics, multi-omics, functional imaging, and artificial intelligence could transform zebrafish from a screening organism into an integrated platform for precision psychiatry and personalized neuropsychopharmacological drug discovery, and improving translation across experimental scales. Zebrafish are best positioned as an efficient intermediate translational model that accelerates mechanism-based anxiolytic discovery while complementing, rather than replacing, mammalian preclinical studies.

Matched MeSH terms: High-Throughput Screening Assays
Fulltext A virtual screening approach for identifying plants with anti H5N1 neuraminidase activity

Ikram NK, Durrant JD, Muchtaridi M, Zalaludin AS, Purwitasari N, Mohamed N, et al.

J Chem Inf Model, 2015 Feb 23;55(2):308-16.
PMID: 25555059 DOI: 10.1021/ci500405g

Recent outbreaks of highly pathogenic and occasional drug-resistant influenza strains have highlighted the need to develop novel anti-influenza therapeutics. Here, we report computational and experimental efforts to identify influenza neuraminidase inhibitors from among the 3000 natural compounds in the Malaysian-Plants Natural-Product (NADI) database. These 3000 compounds were first docked into the neuraminidase active site. The five plants with the largest number of top predicted ligands were selected for experimental evaluation. Twelve specific compounds isolated from these five plants were shown to inhibit neuraminidase, including two compounds with IC50 values less than 92 μM. Furthermore, four of the 12 isolated compounds had also been identified in the top 100 compounds from the virtual screen. Together, these results suggest an effective new approach for identifying bioactive plant species that will further the identification of new pharmacologically active compounds from diverse natural-product resources.

Matched MeSH terms: High-Throughput Screening Assays/methods*
Fulltext Sensitive Detection of Plasmodium vivax Using a High-Throughput, Colourimetric Loop Mediated Isothermal Amplification (HtLAMP) Platform: A Potential Novel Tool for Malaria Elimination

Britton S, Cheng Q, Grigg MJ, Poole CB, Pasay C, William T, et al.

PLoS Negl Trop Dis, 2016 Feb;10(2):e0004443.
PMID: 26870958 DOI: 10.1371/journal.pntd.0004443

INTRODUCTION: Plasmodium vivax malaria has a wide geographic distribution and poses challenges to malaria elimination that are likely to be greater than those of P. falciparum. Diagnostic tools for P. vivax infection in non-reference laboratory settings are limited to microscopy and rapid diagnostic tests but these are unreliable at low parasitemia. The development and validation of a high-throughput and sensitive assay for P. vivax is a priority.
METHODS: A high-throughput LAMP assay targeting a P. vivax mitochondrial gene and deploying colorimetric detection in a 96-well plate format was developed and evaluated in the laboratory. Diagnostic accuracy was compared against microscopy, antigen detection tests and PCR and validated in samples from malaria patients and community controls in a district hospital setting in Sabah, Malaysia.
RESULTS: The high throughput LAMP-P. vivax assay (HtLAMP-Pv) performed with an estimated limit of detection of 1.4 parasites/ μL. Assay primers demonstrated cross-reactivity with P. knowlesi but not with other Plasmodium spp. Field testing of HtLAMP-Pv was conducted using 149 samples from symptomatic malaria patients (64 P. vivax, 17 P. falciparum, 56 P. knowlesi, 7 P. malariae, 1 mixed P. knowlesi/P. vivax, with 4 excluded). When compared against multiplex PCR, HtLAMP-Pv demonstrated a sensitivity for P. vivax of 95% (95% CI 87-99%); 61/64), and specificity of 100% (95% CI 86-100%); 25/25) when P. knowlesi samples were excluded. HtLAMP-Pv testing of 112 samples from asymptomatic community controls, 7 of which had submicroscopic P. vivax infections by PCR, showed a sensitivity of 71% (95% CI 29-96%; 5/7) and specificity of 93% (95% CI87-97%; 98/105).
CONCLUSION: This novel HtLAMP-P. vivax assay has the potential to be a useful field applicable molecular diagnostic test for P. vivax infection in elimination settings.

Matched MeSH terms: High-Throughput Screening Assays/methods*
Computational Analysis and In silico Predictive Modeling for Inhibitors of PhoP Regulon in S. typhi on High-Throughput Screening Bioassay Dataset

Kaur H, Ahmad M, Scaria V

Interdiscip Sci, 2016 Mar;8(1):95-101.
PMID: 26298582 DOI: 10.1007/s12539-015-0273-x

There is emergence of multidrug-resistant Salmonella enterica serotype typhi in pandemic proportions throughout the world, and therefore, there is a necessity to speed up the discovery of novel molecules having different modes of action and also less influenced by the resistance formation that would be used as drug for the treatment of salmonellosis particularly typhoid fever. The PhoP regulon is well studied and has now been shown to be a critical regulator of number of gene expressions which are required for intracellular survival of S. enterica and pathophysiology of disease like typhoid. The evident roles of two-component PhoP-/PhoQ-regulated products in salmonella virulence have motivated attempts to target them therapeutically. Although the discovery process of biologically active compounds for the treatment of typhoid relies on hit-finding procedure, using high-throughput screening technology alone is very expensive, as well as time consuming when performed on large scales. With the recent advancement in combinatorial chemistry and contemporary technique for compounds synthesis, there are more and more compounds available which give ample growth of diverse compound library, but the time and endeavor required to screen these unfocused massive and diverse library have been slightly reduced in the past years. Hence, there is demand to improve the high-quality hits and success rate for high-throughput screening that required focused and biased compound library toward the particular target. Therefore, we still need an advantageous and expedient method to prioritize the molecules that will be utilized for biological screens, which saves time and is also inexpensive. In this concept, in silico methods like machine learning are widely applicable technique used to build computational model for high-throughput virtual screens to prioritize molecules for advance study. Furthermore, in computational analysis, we extended our study to identify the common enriched structural entities among the biologically active compound toward finding out the privileged scaffold.

Matched MeSH terms: High-Throughput Screening Assays/methods*
Fulltext A simple, high-throughput, colourimetric, field applicable loop-mediated isothermal amplification (HtLAMP) assay for malaria elimination

Britton S, Cheng Q, Sutherland CJ, McCarthy JS

Malar J, 2015;14:335.
PMID: 26315027 DOI: 10.1186/s12936-015-0848-3

To detect all malaria infections in elimination settings sensitive, high throughput and field deployable diagnostic tools are required. Loop-mediated isothermal amplification (LAMP) represents a possible field-applicable molecular diagnostic tool. However, current LAMP platforms are limited by their capacity for high throughput.

Matched MeSH terms: High-Throughput Screening Assays/methods*
Approaches for targeted proteomics and its potential applications in neuroscience

Sethi S, Chourasia D, Parhar IS

J Biosci, 2015 Sep;40(3):607-27.
PMID: 26333406

An extensive guide on practicable and significant quantitative proteomic approaches in neuroscience research is important not only because of the existing overwhelming limitations but also for gaining valuable understanding into brain function and deciphering proteomics from the workbench to the bedside. Early methodologies to understand the functioning of biological systems are now improving with high-throughput technologies, which allow analysis of various samples concurrently, or of thousand of analytes in a particular sample. Quantitative proteomic approaches include both gel-based and non-gel-based methods that can be further divided into different labelling approaches. This review will emphasize the role of existing technologies, their advantages and disadvantages, as well as their applications in neuroscience. This review will also discuss advanced approaches for targeted proteomics using isotope-coded affinity tag (ICAT) coupled with laser capture microdissection (LCM) followed by liquid chromatography tandem mass spectrometric (LC-MS/MS) analysis. This technology can further be extended to single cell proteomics in other areas of biological sciences and can be combined with other 'omics' approaches to reveal the mechanism of a cellular alterations. This approach may lead to further investigation in basic biology, disease analysis and surveillance, as well as drug discovery. Although numerous challenges still exist, we are confident that this approach will increase the understanding of pathological mechanisms involved in neuroendocrinology, neuropsychiatric and neurodegenerative disorders by delivering protein biomarker signatures for brain dysfunction.

Matched MeSH terms: High-Throughput Screening Assays/methods

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