Development And Characterization Of Aptamers For The Use In Surface Plasmon Resonance Sensors For The Detection Of Glycated Blood Proteins

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Development and Characterization of Aptamers for the Use in Surface Plasmon Resonance Sensors for the Detection of Glycated Blood Proteins

Author : Nathan George Frederick Reaver
Publisher :
ISBN 13 :
Total Pages : 252 pages
Book Rating : 4.:/5 (867 download)

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Book Synopsis Development and Characterization of Aptamers for the Use in Surface Plasmon Resonance Sensors for the Detection of Glycated Blood Proteins by : Nathan George Frederick Reaver

Download or read book Development and Characterization of Aptamers for the Use in Surface Plasmon Resonance Sensors for the Detection of Glycated Blood Proteins written by Nathan George Frederick Reaver and published by . This book was released on 2013 with total page 252 pages. Available in PDF, EPUB and Kindle. Book excerpt: The concentration ratio of glycated to non-glycated forms of various blood proteins can be used as a diagnostic measure in diabetes to determine a history of glycemic compliance. Depending on a protein's half-life in blood, compliance can be assessed from a few days to several months in the past, which can be used to provide additional therapeutic guidance. The most commonly studied glycated protein for assessing glycemic compliance is glycated hemoglobin. Current glycated protein concentration detection methods are limited in their ability to measure multiple proteins, and are susceptible to interference from other blood pathologies. In this study, DNA aptamers were developed and characterized for use in Surface Plasmon Resonance (SPR) sensors to assess the percentage of hemoglobin that is glycated in a patient. The aptamers were developed by way of a modified Systematic Evolution of Ligands by Exponential Enrichment (SELEX) process which selects DNA sequences that have a high binding affinity to a specific protein. DNA products resulting from this process were sequenced, and identified aptamers were synthesized. The SELEX process was performed multiple times. One process produced aptamers specific to a glycated form of hemoglobin, another produced aptamers specific to non-glycated hemoglobin and aptamers specific to all forms of hemoglobin. A final SELEX process produced aptamers specific to fibrinogen. Equilibrium dissociation constants for the affinity of the identified aptamers to glycated hemoglobin, hemoglobin, and fibrinogen were calculated from fitting a Langmuir binding model to experimental, real time, binding data obtained through SPR. It was determined that two aptamers, GHA1 and GHA2, were selective to glycated hemoglobin, with equilibrium dissociation constants of 11.5 nM and 51.8 nM, respectively; one aptamer, HA2, was selective to non-glycated hemoglobin, with an equilibrium dissociation constant of 187 nM; and one aptamer was selective to both glycated and non-glycated hemoglobin, with equilibrium dissociation constants for the two proteins of 36.9 nM and 35.2 nM, respectively. It was also demonstrated how these aptamers could be used in an SPR sensor. The aptamers identified in this study can potentially be used in a SPR aptamer-based biosensor for detection of glycated hemoglobin, a technology that has the potential to deliver low-cost and immediate glycemic compliance assessment in either a clinical or home setting.

The Development of an Aptamer-based Surface Plasmon Resonance (SPR) Sensor for the Real-time Detection of Glycated Protein

Author : Rui Zheng
Publisher :
ISBN 13 :
Total Pages : 187 pages
Book Rating : 4.:/5 (863 download)

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Book Synopsis The Development of an Aptamer-based Surface Plasmon Resonance (SPR) Sensor for the Real-time Detection of Glycated Protein by : Rui Zheng

Download or read book The Development of an Aptamer-based Surface Plasmon Resonance (SPR) Sensor for the Real-time Detection of Glycated Protein written by Rui Zheng and published by . This book was released on 2012 with total page 187 pages. Available in PDF, EPUB and Kindle. Book excerpt: The direct detection of blood proteins can benefit a number of scientific and clinical applications, one of which is glycated protein monitoring for diabetes diagnosis and treatment. The formation of glycated proteins is proportional to the average glucose concentration over a certain past time period. Thus, the protein glycation level can reflect the long-term glycemic compliance according to a specific protein's half-life. Albumin has a short half-life of 2-3 weeks and the glycated albumin (GA) level could indicate the average plasma glucose concentration on a monthly basis, which provides valuable complementary information for more complete assessment of glycemic control in comparison to conventional glucose and HbA1c testing. (HbA1c is a common long-term glucose indicator with a half-life of 7-8 weeks). According to a literature review, no glycated protein monitor, other than HbA1c, is currently on the commercial market for diabetes guidance. A cost-effective, real-time monitoring approach to assess the HbA1c and other proteins (such as GA) could bring significant advantages for diabetes diagnosis and treatment, whereas the existing monitoring technology does not make this economical. In this research project, the feasibility of using an aptamer-based surface plasmon resonance (SPR) sensors for the real-time detection of glycated blood proteins (i.e., GA) has been studied. For the purpose of establishment, the well documented thrombin binding aptamers were immobilized onto a gold sensing surface using a two-step amine coupling method. The experimental results show that the aptamer functionalized SPR is a well suited sensing platform for selective blood protein detection applications. To obtain the specific receptors that enable selective detection, the human serum albumin (HSA) binding aptamer has then been developed using the magnetic beads-based Systematic Evolution of Ligands by EXponential amplification process (MB-SELEX). The experimental protocol of the in vitro selection process have been standardized. The developed aptamers have shown a common pattern in sequence and a similarity in secondary structures. Furthermore, the glycated albumin (GA) binding aptamers have been identified and the sensor performance has been characterized under different binding conditions. The aptamer functionalized sensor has shown a better performance in both sensitivity and selectivity for the detection of GA compared to the well documented phenylboronate monolayer-based sensor, indicating that aptamers are a better receptor choice for GA detection applications. Finally, a novel multi-channel aptamer functionalization sensor and a feedback data processing model have been developed based on the glycated and non-glycated albumin binding aptamers. The experimental results demonstrate that the aptamer-based SPR sensor has the ability to detect GA levels within a 25% error range. This dissertation provides the essential foundation experiments to demonstrate the feasibility of using aptamer-based SPR sensor for the detection of glycated protein. It is expected that the further engineering and improvement of the sensor will eventually lead to a fast, reliable, label-free, and cost-effective glycated protein detection sensor, which may prove useful in the diagnosis and treatment of diabetes.

Aptamers in Biotechnology

Author : Katharina Urmann
Publisher : Springer Nature
ISBN 13 : 3030540618
Total Pages : 217 pages
Book Rating : 4.0/5 (35 download)

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Book Synopsis Aptamers in Biotechnology by : Katharina Urmann

Download or read book Aptamers in Biotechnology written by Katharina Urmann and published by Springer Nature. This book was released on 2020-07-31 with total page 217 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book reviews the development, characterization and applications of aptamers in different areas of biotechnology ranging from therapeutics to diagnostics and protein purification. Hailed as chemical antibodies, these single-stranded nucleic acid receptors were predicted to supersede antibodies in traditional assays, such as ELISA, within a short time. While this has yet to happen, readers will find in this book a deep insight into the progress of aptamer technology and a critical discussion about the limitations that need to be overcome in order to find wider acceptance and use outside of the still relatively small aptamer-community. This book covers all aspects of aptamer generation and application for the aptamer-experienced reader and curious novice alike, with the addition of an industry perspective on the future of aptamer-use in biotechnology.

Biosensors Based on Aptamers and Enzymes

Author : Man Bock Gu
Publisher : Springer
ISBN 13 : 3642541437
Total Pages : 338 pages
Book Rating : 4.6/5 (425 download)

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Book Synopsis Biosensors Based on Aptamers and Enzymes by : Man Bock Gu

Download or read book Biosensors Based on Aptamers and Enzymes written by Man Bock Gu and published by Springer. This book was released on 2014-07-08 with total page 338 pages. Available in PDF, EPUB and Kindle. Book excerpt: Volumes are organized topically and provide a comprehensive discussion of developments in the respective field over the past 3-5 years. The series also discusses new discoveries and applications. Special volumes are dedicated to selected topics which focus on new biotechnological products and new processes for their synthesis and purification. In general, special volumes are edited by well-known guest editors. The series editor and publisher will however always be pleased to receive suggestions and supplementary information. Manuscripts are accepted in English.

Aptasensors for Point-of-Care Diagnostics

Author : Arpana Parihar
Publisher : CRC Press
ISBN 13 : 1003829503
Total Pages : 375 pages
Book Rating : 4.0/5 (38 download)

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Book Synopsis Aptasensors for Point-of-Care Diagnostics by : Arpana Parihar

Download or read book Aptasensors for Point-of-Care Diagnostics written by Arpana Parihar and published by CRC Press. This book was released on 2023-12-18 with total page 375 pages. Available in PDF, EPUB and Kindle. Book excerpt: The design and fabrication of aptamer-based biosensors for point-of-care testing of disease diagnostic and prognostic is an upthrust and interdisciplinary area of research. This book covers the design and development of novel cost-effective aptamer-based biosensors for disease diagnostic and prognostic including an understanding of health care management in terms of timely updates of disease situations in a particular geographical area. It also discusses the scaling up and market trend of aptamer-based devices for extension of research from lab to market, and end users. Features: Explains the design and fabrication of aptasensors-based diagnostic devices. Includes novel approaches and comprehensive technology overview for diagnosis management towards early-stage detection of various biomarkers associated with several health conditions. Demonstrates possible benefits of combined diagnostics and therapeutic possibilities using aptamer-based point-of-care technologies devices. Discusses emerging implications and recent advances of biosensing platforms for designing and developing aptamer-based point-of-care testing (POCT) devices. Explores aptamer-based approach to develop advanced platforms for medical applications and open arena for state-of-the-art future research. This book is aimed at researchers and graduate students in biomedical engineering, diagnostics, nanobiotechnology, and materials science.

Nucleic Acid Aptamers

Author : Günter Mayer
Publisher : Springer Nature
ISBN 13 : 1071626957
Total Pages : 283 pages
Book Rating : 4.0/5 (716 download)

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Book Synopsis Nucleic Acid Aptamers by : Günter Mayer

Download or read book Nucleic Acid Aptamers written by Günter Mayer and published by Springer Nature. This book was released on 2022-09-26 with total page 283 pages. Available in PDF, EPUB and Kindle. Book excerpt: This updated book reflects improvements in a variety of techniques used to study the aptamer field. Beginning with a section on selection procedures, the volume continues with methods to characterize aptamers’ interaction and structural properties by biophysical approaches, as well as a variety of applications that have been adapted to the aptamer compound class. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Nucleic Acid Aptamers: Selection, Characterization, and Application, Second Edition serves as an ideal guide for researchers aiming to further our understanding of aptamer biology and more.

Aptamers for Medical Applications

Author : Yiyang Dong
Publisher : Springer Nature
ISBN 13 : 9813348380
Total Pages : 470 pages
Book Rating : 4.8/5 (133 download)

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Book Synopsis Aptamers for Medical Applications by : Yiyang Dong

Download or read book Aptamers for Medical Applications written by Yiyang Dong and published by Springer Nature. This book was released on 2021-03-25 with total page 470 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book outlines comprehensively the main medical uses of aptamers, from diagnosis to therapeutics in fourteen chapters. Pioneering topics covered include aptamer pharmaceuticals, aptamers for malign tumors, aptamers for personalized therapeutics and aptamers for point-of-care testing. The book offers an essential guide for medical scientists interested in developing aptamer-based schemes for better theranostics. It is therefore of interest for not only academic researchers, but also practitioners and medical researchers in various fields of medical science, medical research and bio-analytical chemistry.

Aptamers for Analytical Applications

Author : Yiyang Dong
Publisher : John Wiley & Sons
ISBN 13 : 3527342672
Total Pages : 426 pages
Book Rating : 4.5/5 (273 download)

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Book Synopsis Aptamers for Analytical Applications by : Yiyang Dong

Download or read book Aptamers for Analytical Applications written by Yiyang Dong and published by John Wiley & Sons. This book was released on 2019-02-11 with total page 426 pages. Available in PDF, EPUB and Kindle. Book excerpt: An essential guide that puts the focus on method developments and applications in aptamers In recent years, aptamer-based systems have been developed for a wide-range of analytical and medical applications. Aptamers for Analytical Applications offers an introduction to the topic, outlines the common protocols for aptamer synthesis, as well as providing information on the different optimization strategies that can obtain higher affinities to target molecules. The contributors?noted experts on the topic?provide an in-depth review of the characterization of aptamer-target molecule interaction and immobilization strategies and discuss the developments of methods for all the relevant applications. The book outlines different schemes to efficiently immobilize aptamers on substrates as well as summarizing the characterization methods for aptamer-ligand complexes. In addition, aptamer-based colorimetric, enzyme-linked, fluorescent, electrochemical, lateral flow and non-labeling analytical methods are presented. The book also reflects state-of-the-art and emerging applications of aptamer-based methods. This important resource: -Provides a guide to aptamers which provide highly specific and sensitive molecular recognition, with affinities in the range of antibodies and are much cheaper to produce -Offers a discussion of the analytical method developments and improvements with established systems and beyond -Offers a comprehensive guide to all the relevant application areas -Presents an authoritative book from contributors who are noted experts in the field Written for analytical chemists, biochemists, analytical researchers, Aptamers for Analytical Applications is a comprehensive book that adopts a methodological point of view to the important aspects of aptamer generation and modification with a strong emphasis on method developments for relevant applications.

Aptamer-based Sensors

Author : David Andrew Douglas Blair
Publisher :
ISBN 13 : 9780494520055
Total Pages : 248 pages
Book Rating : 4.5/5 (2 download)

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Book Synopsis Aptamer-based Sensors by : David Andrew Douglas Blair

Download or read book Aptamer-based Sensors written by David Andrew Douglas Blair and published by . This book was released on 2008 with total page 248 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Aptamers in Bioanalysis

Author : M. Mascini
Publisher : John Wiley & Sons
ISBN 13 : 0470380764
Total Pages : 346 pages
Book Rating : 4.4/5 (73 download)

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Book Synopsis Aptamers in Bioanalysis by : M. Mascini

Download or read book Aptamers in Bioanalysis written by M. Mascini and published by John Wiley & Sons. This book was released on 2009-04-27 with total page 346 pages. Available in PDF, EPUB and Kindle. Book excerpt: This is the first book to detail bioanalytical technologies and methods that have been developed using aptamers in analytical, medical, environmental, and food science applications. After an introduction to aptamers, aptamer targets, and their general uses, it discusses different applications with particular attention to the comparison between aptamer-based biosensors and methods versus the corresponding immunosensors. Examples of aptamer-based diagnostic techniques include whole-cell protein profiling (proteomics) and medical diagnostics for the distinction of diseased versus healthy states. This is a core reference for analytical chemists, electrochemists, pharmaceutical/medicinal chemists, biotechnologists, and others.

Fabrication of a Surface Plasmon Resonance Platform for the Development of an Electrokintic Surface Plasmon Resonance (EK-SPR) Biosensor

Download Fabrication of a Surface Plasmon Resonance Platform for the Development of an Electrokintic Surface Plasmon Resonance (EK-SPR) Biosensor PDF Online Free

Author : Ornella Sathoud
Publisher :
ISBN 13 : 9780438932159
Total Pages : 98 pages
Book Rating : 4.9/5 (321 download)

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Book Synopsis Fabrication of a Surface Plasmon Resonance Platform for the Development of an Electrokintic Surface Plasmon Resonance (EK-SPR) Biosensor by : Ornella Sathoud

Download or read book Fabrication of a Surface Plasmon Resonance Platform for the Development of an Electrokintic Surface Plasmon Resonance (EK-SPR) Biosensor written by Ornella Sathoud and published by . This book was released on 2019 with total page 98 pages. Available in PDF, EPUB and Kindle. Book excerpt: Point-of-Care Testing (POCT) is a clinical analysis performed near the patient, rather than in a remote, dedicated laboratory. POCT is typically performed by non-laboratory personnel and the results are used for clinical decision-making. Acute Coronary syndrome (ACS) is a group of conditions that considerably decreases or stops blood flow to the heart muscle, causing the heart to be damaged or stressed. Such events cause the release of biomarkers into the blood stream. Myocardial Infarction (MI), commonly called "heart attack", is a well-known ACS that is diagnosed by measuring the change in the level of cardiac biomarkers (preferably troponin) in the blood. This dissertation presents the fabrication of a platform to be adapted as a point-of care sensor with surface plasmon resonance as the detection method. 4-aminophenylalanine is electrografted on the gold sensing surface as the linker to allow receptors immobilization on the sensing surface. Electrokinetic method is to be combined with surface plasmon resonance, to enhance the sensitivity and selectivity of a cardiac protein biosensor. Surface Plasmon Resonance (SPR) is a real time, label free optical method that measures the refractive index change when the analyte binds receptors immobilized at the surface of a metal, here gold. In this report details on how electrografting of 4-aminophenylalanine on gold thin film permits the fabrication of an SPR biosensor platform that could possibly be as a troponin biosensor. An overview on the sensing surface characterization to confirm surface modification is offered as well as the sensor performance using bovine serum albumin bovine serum albumin antibodies.

Aptasensors for Food Safety

Author : Raju Khan
Publisher : CRC Press
ISBN 13 : 1040118607
Total Pages : 547 pages
Book Rating : 4.0/5 (41 download)

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Book Synopsis Aptasensors for Food Safety by : Raju Khan

Download or read book Aptasensors for Food Safety written by Raju Khan and published by CRC Press. This book was released on 2024-10-04 with total page 547 pages. Available in PDF, EPUB and Kindle. Book excerpt: This reference book provides updated knowledge on fundamentals and applications of aptasensors in food science. It discusses aptamer selection strategies including Systematic Evolution of Ligands through EXponential enrichment (SELEX), Capture SELEX, Cell SELEX, and GO SELEX strategies for aptasensor design involving well-studied and newly emerged nanomaterials, food analysis focusing on its safety, aptasensing techniques relevant to detection of food pathogens, pesticides, veterinary drug residues, toxins, heavy metals, allergens, and future perspectives of aptamers in food science and technology. The book focuses on recent developments in aptasensors and their use in detection of contaminants in food. The book is an excellent resource for R&D organizations dedicated to the development of biosensors in general and aptasensors in particular for the detection of environmental contaminants in the food supply chain. Also, the contents of the book will be useful to researchers, postgraduate students, analytical chemists, food scientists, and regulatory authorities.

Aptamers as Biosensors

Author : Colette Dalton
Publisher :
ISBN 13 :
Total Pages : 0 pages
Book Rating : 4.:/5 (141 download)

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Book Synopsis Aptamers as Biosensors by : Colette Dalton

Download or read book Aptamers as Biosensors written by Colette Dalton and published by . This book was released on 2010 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Aptamers are short, single strands of artificial nucleic acids which bind to specific targets. This binding is similar to the binding exhibited by antibodies but aptamers offer improved properties and consequently are replacing antibodies in some applications. This first part of this project attempted to generate an aptamer which would bind to the molecules amphetamine and methamphetamine, with high selectivity and affinity. The process for generating an aptamer is called Systematic Evolution of Ligands by Exponential Enrichment (SELEX) and is carried out in vitro. The strength of an aptamer / target binding interaction can be assessed using a variety of different techniques and in this work an Enzyme Linked Immunosorbent Assay (ELISA) was investigated. Other methods including surface plasmon resonance (SPR) and circular dichroism were attempted. The phenomenon of naturally occurring RNA molecules with catalytic properties (ribozymes) were the inspiration for the development of catalytic aptamers. Consequently, a novel SELEX system was deigned to select aptamers capable of catalysing a Diels Alder reaction between cyclohexadiene modified DNA and maleimide. A number of potential sequences were isolated and when one such sequence was incorporated into a test reaction a cycloadduct resulting from a successful Diels-Alder reaction was confirmed using MALDI - MS. In addition to generating novel aptamers, this thesis details work on the application of existing aptamers with the aim of creating novel assays and expanding the potential use of aptamers. Two protein targets, namely Protein kinase C (PKC) and thrombin were chosen and their respective aptamers investigated for use in metallic nanoparticle conjugates. The combination of aptamers and nanoparticles (in particular metallic nanoparticles) has grown in prominence in the literature and is regarded as an exciting development due to their desirable optical properties. Consequently, novel assays based on UV-visible spectroscopy were developed using the two specified DNA aptamers conjugated to gold and / or silver nanoparticles. In addition, the possibility of incorporating Surface enhanced resonance Raman scattering (SERRS) analysis was investigated.

Aptamers for Parallel Measurements of Cellular Molecules

Author : Shengnan Xie
Publisher :
ISBN 13 :
Total Pages : 366 pages
Book Rating : 4.3/5 (129 download)

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Book Synopsis Aptamers for Parallel Measurements of Cellular Molecules by : Shengnan Xie

Download or read book Aptamers for Parallel Measurements of Cellular Molecules written by Shengnan Xie and published by . This book was released on 2010 with total page 366 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Study of Aptamer Selection Methodologies for Developing Piezoelectric Quartz Crystal Biosensors to Detect Albumin in Urine, Malaria and Sars Protein Biomarkers in Serum

Download Study of Aptamer Selection Methodologies for Developing Piezoelectric Quartz Crystal Biosensors to Detect Albumin in Urine, Malaria and Sars Protein Biomarkers in Serum PDF Online Free

Author : Dharmatov Rahula Barlongo Albano
Publisher :
ISBN 13 : 9781361337363
Total Pages : pages
Book Rating : 4.3/5 (373 download)

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Book Synopsis Study of Aptamer Selection Methodologies for Developing Piezoelectric Quartz Crystal Biosensors to Detect Albumin in Urine, Malaria and Sars Protein Biomarkers in Serum by : Dharmatov Rahula Barlongo Albano

Download or read book Study of Aptamer Selection Methodologies for Developing Piezoelectric Quartz Crystal Biosensors to Detect Albumin in Urine, Malaria and Sars Protein Biomarkers in Serum written by Dharmatov Rahula Barlongo Albano and published by . This book was released on 2017-01-26 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: This dissertation, "Study of Aptamer Selection Methodologies for Developing Piezoelectric Quartz Crystal Biosensors to Detect Albumin in Urine, Malaria and SARS Protein Biomarkers in Serum" by Dharmatov Rahula Barlongo, Albano, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: Due to the high affinity and selectivity of aptamer selected from DNA library towards target proteins, the use of specific protein biomarkers to assist early diagnosis, and the recent rapid development of piezoelectric quartz crystal (PQC) biosensor technology, the research reported is focused on developing PQC biosensor using aptamers selected from a DNA library as selective coating for the detection of albumin in urine, two protein biomarkers for malaria in sera, and the determination of trace amount of SCV helicase protein from SAR in sera. Three new aptamer-PQC biosensors are developed with research conducted on aptamer selection, optimization of coating methods for aptamers on PQC, and coupling paramagnetic nanoparticle technology with PQC biosensor in flow injection analysis (FIA) to enrich analyte and eliminate sample matrix interference. A new approach for aptamer selection is successfully attempted in the development of the first aptamer-PQC biosensor with an aim to offer an early detection of microalbuminuria enabled by an increase in detection sensitivity with aptamer selected by its actual binding with albumin-PQC biosensor. The aptamer-PQC sensor developed has shown to give a working range between 0.1 to 10 ug/mL, a detection limit (S/N = 3, n=3) of 0.048 μg/ml, repeatability of RSD = 6.8% (n=3), a response time of 1 minute and a throughput of 60 samples/hour. The developed piezoelectric aptamer sensor is shown to have sufficient sensitivity to detect microgram quantities of albumin in urine. Aptamer-PQC biosensors are developed for the detection of two protein biomarkers, PfLDH for general diagnosis and PfHRP-2 for a specific diagnosis of Plasmodium falciparum malaria parasitemia, in sera. The results show satisfactory working range from 10-100 ng/mL for both PfLDH and PfHRP-2 protein biomarker, low detection limit of 1.8 and 4.7 ng/mL and satisfactory repeatability (%RSD, n = 3) of 7.4% and 9.2% for PfHRP-2 and PfLDH respectively. It is first report for aptamer based PQC biosensor to detect malarial PfHRP-2 and PfLDH at ppb range to meet the requirement for their diagnosis. To meet the highly demanding challenge for detecting protein biomarkers at pg/mL level in high-protein sera sample, the coupling of paramagnetic nanoparticle technology with aptamer-PQC biosensor was attempted for developing aptamer-PQC biosensor for selective detection of SCV helicase protein produced from SARS CoV replication. The coupling of aptamer coated paramagnetic nanoparticles for sample pretreatment to aptamer-PQC biosensor has shown to detect helicase protein in one-minute assay with a detection limit of 350 pg/mL. The aptamer-coated crystal exhibits a frequency shift linearly proportional to the concentration of SARS helicase from 1 to 1000 ng/mL with a correlation coefficient of 0.9975 and a repeatability of 6.8% (%RSD, n=3). After the enrichment procedure, recoveries of 102% and 119% were achieved using samples spiked with SARS helicase at concentrations of 10 ng/mL and 1.0 ng/mL respectively. It is the first report to detect for SCV helicase protein using PQC biosensor at pg/mL level after magnetic bead enrichment. DOI: 10.5353/th_b5185935 Subjects: Biochemical markers Albumins Piezoelectric polymer biosensors

Development and Characterization of High-throughput Methods and Technologies for in Vitro RNA Aptamer Selections

Author : Kylan Szeto
Publisher :
ISBN 13 :
Total Pages : 197 pages
Book Rating : 4.:/5 (892 download)

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Book Synopsis Development and Characterization of High-throughput Methods and Technologies for in Vitro RNA Aptamer Selections by : Kylan Szeto

Download or read book Development and Characterization of High-throughput Methods and Technologies for in Vitro RNA Aptamer Selections written by Kylan Szeto and published by . This book was released on 2014 with total page 197 pages. Available in PDF, EPUB and Kindle. Book excerpt: Aptamers are an emerging class of molecules that are valued for their ability to bind with high affinity and specificity to a desired target. These DNA or RNA ligands can be synthesized easily in the lab making them much more stable, reproducible, and cost-effective than other affinity reagents such as antibodies. These molecules possess a diverse and versatile set of abilities upon binding, making them useful not only in biotechnology and diagnostics, but also in therapeutics. Aptamers are typically generated through an in vitro process called SELEX (Systematic Evolution of Ligands by EXponential Enrichment). SELEX is an iterative process whereby a library of random sequences is exposed to a target, and bound sequences are selected and amplified making a pool enriched for improved binding over the original library. This process can be repeated until the best binding sequence dominates the pool. In this way, aptamers can be generated to nearly any conceivable target or molecule, overcoming the immunological limitations in generating antibodies. By manipulating the conditions within this selection process, aptamers can also be generated to bind in non-physiological environments, giving them the ability to perform in diverse applications. To help accelerate the discovery of aptamers, we developed a modular microcolumn technology that decreases reagent consumption, while permitting multiplex SELEX, allowing for more sophisticated selection schemes. By characterizing aptamer enrichments for all the available parameters, we have significantly increased the selection efficiency, while illustrating the failures of simple binding theories to non-classical modes of selection. We have also demonstrated the power of high-throughput sequencing for early identification of aptamers before they fully converge. This was used to validate a new method which optimizes selection time over individual selection cycle efficiencies. Surprisingly, our results also show the new method significantly improves selection efficiency, bringing into question some common beliefs derived from SELEX theory. Finally, we scaled our microcolumn technology to an automatable 96-well microplate format, and demonstrated its utility by characterizing and validating specific, non-specific, and background binding behavior of several RNA aptamers. The results reveal binding behaviors that fundamentally limit the performance and sensitivity of aptamer selections.

Aptamers for Food Applications

Author : Yiyang Dong
Publisher : Elsevier
ISBN 13 : 0323919030
Total Pages : 350 pages
Book Rating : 4.3/5 (239 download)

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Book Synopsis Aptamers for Food Applications by : Yiyang Dong

Download or read book Aptamers for Food Applications written by Yiyang Dong and published by Elsevier. This book was released on 2023-06 with total page 350 pages. Available in PDF, EPUB and Kindle. Book excerpt: Aptamers for Food Applications: Safety, Authenticity, and Integrity outlines the main applications of aptamers to ensure food safety, quality and compliance that are crucial for food science, public health and human sustainability. Aptamers or aptamer-based analytical devices are comprehensively summarized to tackle various food contamination or food adulteration problems. Topics covered in this book include aptamers for foodborne microbial pathogens, pesticide or veterinary drug residues, heavy metals, biotoxins, abused food additives, illegal food adulterants, POPs, GMOs, food allergens and food contact material migrants. It is therefore of interest for not only analytical investigators and food safety inspectors, but also regulatory practitioners and risk assessors in various food science and research fields.