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Liquid biopsy is a technology that exhibits potential to detect cancer early,monitor therapies,and predict cancer prognosis due to its unique characteristics,including noninvasive sampling and real-time analysis.Circulating tumor cells(CTCs)and extracellular vesicles(EVs)are two important components of circu-lating targets,carrying substantial disease-related molecular information and playing a key role in liquid biopsy.Aptamers are single-stranded oligonucleotides with superior affinity and specificity,and they can bind to targets by folding into unique tertiary structures.Aptamer-based microfluidic platforms offer new ways to enhance the purity and capture efficiency of CTCs and EVs by combining the advantages of microfluidic chips as isolation platforms and aptamers as recognition tools.In this review,we first briefly introduce some new strategies for aptamer discovery based on traditional and aptamer-based micro-fluidic approaches.Then,we subsequently summarize the progress of aptamer-based microfluidics for CTC and EV detection.Finally,we offer an outlook on the future directional challenges of aptamer-based microfluidics for circulating targets in clinical applications.
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Polymer nanomaterials have been attracted more and more attention because of their advantages such as long circulation, reduced immunogenicity and less side effects, and have become a hot research topic in nanomaterials. However, the number of polymer nanomedicines successfully applied in clinical application is very limited, and the unsatisfactory pharmacokinetic behavior is one of the main reasons for thisresult. After polymer nanoparticles enter the body, they will release free drugs and polymer excipients. Polymer nanoparticles are the loaded drugs and free drugs are the active chemicals for efficacy, while polymer excipients may cause excipient drug interactions. Therefore, the focus of the pharmacokinetics study of polymer nanoparticles should not be only limited to the free drugs themselves, but should also focus on the loaded drugs, free drugs and polymer excipients. The dynamic changes of polymer excipients and their metabolites pose new requirements and challenges for the bioanalysis of polymer nanomedicines. The characteristics and application scope of common analytical methods for detection polymer nanomedicines including chromatographic assay will be discussed in this paper. Moreover, this review will also summarize the absorption, distribution, metabolism and excretion of polymer nanomedicines. We hope this review will provide reference for the pharmacokinetics study, safety and effectiveness evaluation of polymer nanomedicines.
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Sample preparation is considered as the bottleneck step in bioanalysis because each biological matrix has its own unique challenges and complexity.Competent sample preparation to extract the desired analytes and remove redundant components is a crucial step in each bioanalytical approach.The matrix effect is a key hurdle in bioanalytical sample preparation,which has gained extensive consideration.Novel sample preparation techniques have advantages over classical techniques in terms of accuracy,automation,ease of sample preparation,storage,and shipment and have become increasingly popular over the past decade.Our objective is to provide a broad outline of current developments in various bioanalytical sample preparation techniques in chromatographic and spectroscopic examinations.In addition,how these techniques have gained considerable attention over the past decade in bioanalytical research is mentioned with preferred examples.Modern trends in bioanalytical sample preparation techniques,including sorbent-based microextraction techniques,are primarily emphasized.
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GeXIVA[1,2] is a new type of conotoxin recently discovered in the transcriptome of Conus generalis and it is expected to be used clinically as a new type of analgesic. This study established and verified a sandwich enzyme-linked immunosorbent assay method for the marine drug GeXIVA[1,2] in the plasma of rats and Beagle dogs. The mouse monoclonal antibody 4B2 and biotin-labeled rabbit polyclonal antibody 2# were developed. The checkerboard method was used to optimize the antibody pairing concentration, minimum dilution ratio, incubation temperature, and incubation time to establish an antibody sandwich ELISA detection method. Verify the established testing methods. The established ELISA method has a quantitative range of 1.25-80 ng·mL-1 in rat and Beagle plasma. The precision, accuracy, selectivity, specificity, stability, dilution linearity, and hook effect all meet the requirements for biological sample analysis. All the procedures for the animal experiments were approved by the Animal Ethics Committee of the Institute (Permit Number: IACUC-DWZX-2020-698). This method can support the preclinical pharmacokinetic study of the marine drug GeXIVA.
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Oligonucleotides have attracted the widespread attention in disease diagnosis and gene therapy. At present, the nucleic acid drugs are at the forefront of biomedical and pharmaceutical research. The bioanalysis of therapeutic oligonucleotides has been slow, however, due to the requirements for pharmacokinetic/toxicokinetic and pharmacodynamic studies in pharmaceutical development. Conventionally, the hybridization-enzyme linked immunosorbent assay (hybridization-ELISA) is widely used in the bioanalysis of therapeutic oligonucleotides. Recentlly, many technologies such as real-time quantitative PCR (qPCR) and high performance liquid chromatography (HPLC)-based technologies have also showed a broad application prospects in the bioanalysis of therapeutic oligonucleotides. However, each technology has its own advantages and limitations. This review summarizes the currently used techniques in the bioanalysis of oligonucleotide therapeutics and reviews the challenges of regulated bioanalysis.
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The new antiepileptic drugs perampanel,retigabine,rufinamide and stiripentol have been recently approved for different epilepsy types.Being them an innovation in the antiepileptics armamentarium,a lot of investigations regarding their pharmacological properties are yet to be performed.Besides,considering their broad anticonvulsant activities,an extension of their therapeutic indications may be worthy of investigation,especially regarding other seizure types as well as other central nervous system disorders.Although different liquid chromatographic (LC) methods coupled with ultraviolet,fluores-cence,mass or tandem-mass spectrometry detection have already been developed for the determination of perampanel,retigabine,rufinamide and stiripentol,new and more cost-effective methods are yet required.Therefore,this review summarizes the main analytical aspects regarding the liquid chro-matographic methods developed for the analysis of perampanel,retigabine (and its main active metabolite),rufinamide and stiripentol in biological samples and pharmaceutical dosage forms.Furthermore,the physicochemical and stability properties of the target compounds will also be addressed.Thus,this review gathers,for the first time,important background information on LC methods that have been developed and applied for the determination of perampanel,retigabine,rufinamide and stiripentol,which should be considered as a starting point if new (bio)analytical techniques are aimed to be imnlemented for these drugs.
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Con el propósito de caracterizar la praxis Bioanalítica desde la matriz epistemológica vigente, la investigación se realizó bajo el Enfoque Integrador Transcomplejo. Para el abordaje de la problemática de estudio se empleó la complementariedad metódica. El grupo humano quedó constituido por cinco docentes Bioanalistas, a los que se le realizó una entrevista focalizada y en profundidad. Entre los hallazgos se evidenció que la praxis bioanalítica fue modelada desde la modernidad como un ejercicio mecanicista, repetitivo, desarticulado de lo social y lo humano. La modernidad es reproducida en las universidades mediante una formación vertical, donde no se articula lo biológico con lo social, con lo histórico o lo cultural; ya que la malla curricular está definida por un grupo de asignaturas cargados de contenidos procedimentales, impartidos a partir de objetivos de forma aislada, que centra la atención en la enfermedad y concibe su praxis desde roles y tareas, dándole mayor énfasis al rol de analista. Desde esta perspectiva, se niega los aportes dados por la epistemología, la historia, lo sociología, antropología y la educación al saber Bioanalítico, quedando limitado su impacto en la Salud Pública. Para redimensionar la concepción social del Bioanálisis y su praxis es necesario que se asuman nuevos paradigmas y nuevas metodologías y la Transcomplejidad constituye una opción para dar respuesta a esta demanda.
In purpose to characterize the Bioanalytical praxis from the current epistemological matrix, the research was carried out under the TranscomplexIntegrative Approach. Methodical complementarity was used to address the study problem. The human group was made up of five Bioanalyst teachers, who were given a focused and in-depth interview. Among the findings, it was evident that bioanalytic praxis was modeled since modernity as a mechanistic, repetitive, disjointed exercise of the social and the human. Modernity is reproduced in universities through vertical training, where the biological is not articulated with the social, with the historical or the cultural; since the curricular mesh is defined by a group of subjects loaded with procedural content, taught from objectives in isolation, which focuses attention on the disease and conceives its praxis from roles and tasks, giving greater emphasis to the role of analyst. From this perspective, the contributions given by epistemology, history, sociology, anthropology and education to Bioanalytical knowledge are denied, leaving their impact on Public Health limited. To resize the social conception of Bioanalysis and its praxis, it is necessary to assume new paradigms and new methodologies and Transcomplexity constitutes an option to respond to this demand.
Subject(s)
Humans , Biological Assay , Chemistry Techniques, Analytical , Public Health , Social Values , Venezuela , Interview , Biomedical Research/methods , Professional TrainingABSTRACT
China's biologic drug research and development is on a fast track lane. The rising cost of new drug development has not improved the success rate of drug launch, and a shift in the model of drug development is urgently needed. In order to improve the success rate, a biomarker strategy of the new drug development model has been proposed and is generally accepted. This paper reviews the research and development of new translational medicine drugs with biomarkers as the core, the application of biomarkers in the clinical study of biological drugs, biomarker biological analysis strategies, and the opportunities and challenges of biomarkers in the clinical study of biological drugs. By comparing international standards, seeking China's advantages and seeking opportunities from the research and development model of translational medicine based on biomarkers, China can make innovative drugs with global influence in the near term.
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During the last decade high-throughput in vitro absorption, distribution, metabolism and excretion (HT-ADME) screening has become an essential part of any drug discovery effort of synthetic molecules. The conduct of HT-ADME screening has been"industrialized"due to the extensive development of software and automation tools in cell culture, assay incubation, sample analysis and data analysis. The HT-ADME assay portfolio continues to expand in emerging areas such as drug-transporter interactions, early soft spot identification, and ADME screening of peptide drug candidates. Additionally, thanks to the very large and high-quality HT-ADME data sets available in many biopharma companies, in silico prediction of ADME properties using machine learning has also gained much momentum in recent years. In this re-view, we discuss the current state-of-the-art practices in HT-ADME screening including assay portfolio, assay automation, sample analysis, data processing, and prediction model building. In addition, we also offer perspectives in future development of this exciting field.
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Formulation/pharmaceutical excipients play a major role in formulating drug candidates, with the ob-jectives of ease of administration, targeted delivery and complete availability. Many excipients used in pharmaceutical formulations are orphanized in preclinical drug discovery. These orphan excipients could enhance formulatability of highly lipophilic compounds. Additionally, they are safe in preclinical species when used below the LD50 values. However, when the excipients are used in formulating compounds with diverse physico-chemical properties, they pose challenges by modulating study results through their bioanalytical matrix effects. Excipients invariably present in study samples and not in the cali-bration curve standards cause over-/under- estimation of exposures. Thus, the mechanism by which excipients cause matrix effects and strategies to nullify these effects needs to be revisited. Furthermore, formulation excipients cause drug interactions by moderating the pathways of drug metabolizing en-zymes and drug transport proteins. Although it is not possible to get rid of excipient driven interactions, it is always advised to be aware of these interactions and apply the knowledge to draw meaningful conclusions from study results. In this review, we will comprehensively discuss a) orphan excipients that have wider applications in preclinical formulations, b) bioanalytical matrix effects and possible ap-proaches to mitigating these effects, and c) excipient driven drug interactions and strategies to alleviate the impacts of drug interactions.
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Liquid chromatography-tandem mass spectrometry (LC-MS) is a promising alternative or complementary method for traditional ligand-binding assays (LBA) in antibody drug bioanalysis. However, issues related to method development, sample preparation, sensitivity and quantitative accuracy need to be addressed. This paper reviews progress in bioanalysis of antibody drugs by LC-MS methods, introduces the principle of the LC-MS method for the analysis of antibody drugs, and describes the challenges faced in quantitative antibody analysis by the LC-MS method. New strategies that can be used to deal with these challenges include: selection of surrogate peptides, purification and enrichment of samples, improvement in enzymatic digest efficiency, enrichment of peptides, and use of low rate LC. We review the application of LC-MS technology in the biological analysis of antibody drugs and discuss the prospect of using the LC-MS method for the analysis of antibody drugs.
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<p><b>Background</b>Nanotechnology is emerging as a promising tool to perform noninvasive therapy and optical imaging. However, nanomedicine may pose a potential risk of toxicity during in vivo applications. In this study, we aimed to investigate the potential toxicity of rare-earth nanoparticles (RENPs) using mice as models.</p><p><b>Methods</b>We synthesized RENPs through a typical co-precipitation method. Institute of Cancer Research (ICR) mice were randomly divided into seven groups including a control group and six experimental groups (10 mice per group). ICR mice were intravenously injected with bare RENPs at a daily dose of 0, 0.5, 1.0, and 1.5 mg/kg for 7 days. To evaluate the toxicity of these nanoparticles in mice, magnetic resonance imaging (MRI) was performed to assess their uptake in mice. In addition, hematological and biochemical analyses were conducted to evaluate any impairment in the organ functions of ICR mice. The analysis of variance (ANOVA) followed by a one-way ANOVA test was used in this study. A repeated measures' analysis was used to determine any significant differences in white blood cell (WBC), alanine aminotransferase (ALT), and creatinine (CREA) levels at different evaluation times in each group.</p><p><b>Results</b>We demonstrated the successful synthesis of two different sizes (10 nm and 100 nm) of RENPs. Their physical properties were characterized by transmission electron microscopy and a 980 nm laser diode. Results of MRI study revealed the distribution and circulation of the RENPs in the liver. In addition, the hematological analysis found an increase of WBCs to (8.69 ± 0.85) × 10/L at the 28 day, which is indicative of inflammation in the mouse treated with 1.5 mg/kg NaYbF:Er nanoparticles. Furthermore, the biochemical analysis indicated increased levels of ALT ([64.20 ± 15.50] U/L) and CREA ([27.80 ± 3.56] μmol/L) at the 28 day, particularly those injected with 1.5 mg/kg NaYbF:Er nanoparticles. These results suggested the physiological and pathological damage caused by these nanoparticles to the organs and tissues of mice, especially to liver and kidney.</p><p><b>Conclusion</b>The use of bare RENPs may cause possible hepatotoxicity and nephritictoxicity in mice.</p>
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Aptamers are single-stranded oligonucleotides ( DNA or RNA ) selected through a technology termed "Systematic evolution of ligands by exponential enrichment" ( SELEX ) . In addition to high affinity and high specificity for their target molecules, aptamers have some advantages such as low molecular weight, easy synthesis, high chemical stability, low immunogenicity, and convenient modification. Based on the Cell-SELEX technique, a panel of aptamers which can specifically recognize target cell lines has been generated. By targeting specific membrane proteins in their native state, these aptamers can identify subtle molecular differences among different cell lines, thus have attracted a broad interest in biomedical research. In this review, we summarized the development of aptamers and their use in detection, profiling and imaging of tumor cells. Also, their perspectives were discussed.
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In the past ten years, the development of electrochemiluminescent ( ECL ) analytical methods based on various types of nanostructures has become a research hotspot. Nanocluster, an intermediate between molecules and conventional nanoparticles, is renowned for its luminescent feature. The first report on ECL for nanoclusters can be traced back to 2009 . Here we summarized the main research progresses since 2011 . Firstly, the preparation of ECL-related nanoclusters was briefly introduced. Then, the mechanisms and applications of ECL by nanoclusters were described. To improve ECL performances, two main strategies, i. e. , nanostructure-based ECL enhancement and biological signal amplification were proposed. Besides, the nanoclusters as the energy transfer receptors in ECL systems were also discussed. In prospect part, the future development of ECL by nanoclusters was considered. We believed that the synthesis of high quality nanoclusters, the reveal of ECL structure-activity relationships, the rationale design and application of near-infrared ECL, and the role of ECL in the interdisciplinary research were the main problems we faced in the future.
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Teniendo en cuenta la insuficiente bibliografía disponible para los estudiantes de la carrera de bioanálisis clínico, sobre los procedimientos técnicos de avanzada, así como la escasa aplicación de los métodos con tecnología novedosa en los centros asistenciales, se diseñó un hiperentorno de enseñanza-aprendizaje, para que fuera utilizado como material de consulta, preparación y autoevaluación en esta asignatura. Para su confección como plataforma se empleó la herramienta CrheaSoft 2.2 y se estructuró en diferentes módulos: Inicio, Temario, Glosario, Ejercicios, Mediateca, Complemento, Créditos y Ayuda. La aplicación del módulo Ejercicio en un grupo de estudiantes de quinto año de la carrera demostró que el producto constituyó un importante material de apoyo a la docencia en esta especialidad.
Keeping in mind the scarce literature available for the students of the clinical bioanalysis career, on the advanced technical procedures, as well as the scarce use of the methods with novel technology in the assistance institutions, a teaching-learning hyperentorno was designed, so that it was used as advisory, preparing and self-evaluating material in this subject. For making it a platform the tool CrheaSoft 2.2 was used and it was structured in different modules: Beginning, Topics, Glossary, Exercises, Mediateca, Complement, Credits and Helps. The use of the module Exercise in a group of students of fifth year of the career showed that the product constituted an important support material for teaching this specialty.
Subject(s)
Virtual Reality , Education, DistanceABSTRACT
Objective: To develop a sensitive and rapid liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method for the simultaneous determination of wogonin, coptisine, berberine, palmatine, jatrorrhizine, phellodendrine, magnoflorine, and wogonoside in rat plasma and to evaluate the pharmacokinetic characteristics of Huanglian Jiedu Decoction (HJD). Methods: LC separation was performed on an Acquity HSS T3 column (100 mm × 2.1 mm, 1.8 μm) using gradient elution with the mobile phase consisting of acetonitrile and 0.1% formic acid water. The detection was accomplished by using positive electrospray ionization in multiple-reaction monitoring mode. Plasma samples were pretreated by protein precipitation. Results: The method showed a good linearity over a wide concentration range (r2 > 0.99). The lower limits of quantification were 0.20 ng/mL for coptisine and phellodendrine, 0.48 ng/mL for berberine, 0.10 ng/mL for jatrorrhizine, 0.32 ng/mL for magnoflorine, 0.30 ng/mL for palmatine, and 4.80 ng/mL for wogonin and wogonoside, respectively. The intra- and inter-day precision of the analytes was less than 12.11%, while the accuracy was between -14.46% and 4.86%. The mean recovery of all the analytes ranged from 93.10% to 110.91%. Conclusion: This validated method offers the advantages of high sensitivity. It is successfully applied to evaluating the pharmacokinetic properties of HJD. © 2014 Tianjin Press of Chinese Herbal Medicines.
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Se realizó un estudio descriptivo y transversal de los 76 estudiantes del cuarto año de la carrera de bioanálisis clínico en la Facultad de Tecnología de la Salud "Dr. Juan Manuel Páez Inchausti" de Santiago de Cuba, desde septiembre del 2012 hasta enero del 2013, con vistas a evaluar las habilidades prácticas de estos en el procesamiento de muestras con equipos de alta tecnología. El empleo de métodos teóricos y empíricos propios de la investigación pedagógica, reveló insuficiencias en las habilidades prácticas relacionadas con la calibración y manipulación de los equipos, debido a la influencia de diferentes factores. Asimismo, se constató la necesidad de emplear medios alternativos de enseñanza que propicien las vías de solución para dicho problema, y que a su vez sean utilizados como material de consulta, preparación y autoevaluación de los educandos.
A descriptive and cross sectional study of the 76 students of the fourth year of the clinical bioanalysis career in "Dr. Juan Manuel Páez Inchausti" Health Technology Faculty, Santiago de Cubawas carried out from September, 2012 to January, 2013, aimed at evaluating their practical skills in the processing of samples with high-technology equipments. The use of theoretical and empiric methods characteristic of the pedagogic investigation, revealed deficiencies in the practical skills related to the calibration and manipulation of the equipments, due to the influence of different factors. Also, the necessity of using teaching alternative means giving solution to this problem was verified, and that, in turn, could be used as consulting, preparation and self-assessment material for the students.
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Micellar liquid chromatography (MLC) is one of the very important analytical techniques. It contains mobile phase added with surfactants above its critical micellar concentration and the stationary phase is modified with surfactant monomers. So, micelles alter the solubilising capability of the mobile phase which forms diverse interactions with major implications in retention and selectivity. It is an alternative to conventional reversed phase liquid chromatography. It allows direct injection of physiological fluids, analysis of pharmaceutical compounds, physiological partitioning process. MLC (micelle liquid chromatography) has proved time saving as compared to other analytical technique like HPLC and ion-pairing. Applicability of MLC is increased in the field of bioanalysis. It is used to analyze different samples of drugs in serum, urine, food etc. In this review we have focused on the various examples of use of MLC in bioanalysis. This review also contains basic information about MLC such as micellar mobile phase, stationary phase, surfactants, and fundamental studies such as retention behavior and partition coefficient.
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Qualitative and quantitative analyses of biological samples containing drugs, toxicants and endogenous substances play an important role in the researches of life sciences, as well as in new drug discovery and development. Biological samples are characterized by complex matrix, multiple endogenous interferences, significantly lower concentrations of measured analytes compared to endogenous components and small sampling volume. Consequently, it often requires bioanalysis methods with superior specificity, high sensitivity and good reproducibility. The two-dimensional liquid chromatography (2D-LC) technique, which allows for high peak capacity, significant reduced matrix effect and carryover of complex matrix samples and automated sample pre-treatment and analysis, has been the powerful solution to the separation and analysis of biological sample and widely applied to environment, food and pharmaceutical analysis. On the basis of introduction of the principle and equipments of 2D-LC, the application of this technique in the pharmacokinetics, toxicological and biological study was reviewed.
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OBJECITVE: To we summarize the background, evolution and implementation of incurred sample reanalysis (ISR) for bioanalysis.