Day 3 :
- Track:9 Applications of Analytical and Bio analytical Methods
Track:10 Diagnostic Assays and Test Kits
Track:11 New Instrumentation and Equipment
Session Introduction
Kabir Hussain
MedImmune, UK
Title: Immunoassays in drug development – Comparing and contrasting different platforms
Biography:
Kabir Hussain has a BSc degree in Cancer Biology & Immunology from the University of Bristol, UK. As part of the Clinical Pharmacology and DMPK group, he is responsible for the development, validation and execution of Immunoassays used in the analysis of samples from clinical and non-clinical studies, including GLP toxicology studies. He works in a GLP certified laboratory and consequently has experience working in accordance with the OECD principles of Good Laboratory Practice. He is a Principle Investigator for GLP multi-site studies and a Study Director for method validation studies. He previously worked at a CRO developing and validating immunoassays.
Abstract:
The drug development process requires sensitive and robust assays to enable PK/PD and Immunogenicity assessment of biologics. Immunoassays are commonly employed to this purpose, utilising the target-specific properties of antibodies to detect the analyte of interest. With the rapid advances in technology, the range of immunoassay platforms available is growing whilst existing platforms are becoming more refined, enabling bioanalytical scientists to choose a platform that is tailored to their needs. Various factors can influence the choice of platform, these can be either assay related such as sensitivity or assay type (quantitative/immunogenicity), or more general factors such as time and cost effectiveness. The Meso Scale Discovery® platform has become established as a reliable platform, particularly for immunogenicity assessment whereby a homogenous solution phase incubation of the sample with the capture and detection reagents provides a simple, easy to perform assay. The GyrolabTM is a largely automated platform that provides a time effective option allowing quick turnaround for analysis of samples. The CiraTM immunoassay platform from Aushon Biosystems, Inc. provides highly sensitive assays that are ideal for Biomarker analysis and also specialises in multiplexing capabilities, making it an attractive option for pharmacodynamics assessment. Carefully selecting the appropriate platform can improve the likelihood of successfully navigating biologics through the drug development process.
Lihan Tan
Bioprocessing Technology Institute, Singapore
Title: Measurement of histones in mammalian cell culture harvest
Biography:
Lihan Tan has completed her PhD from National University of Singapore, Singapore. Currently, she is working as a research scientist in downstream processing group, Bioprocessing Technology Institute, A*STAR, Singapore. Her research interest is on developing new platforms for protein or virus purifications, and on analytical techniques to characterize the biomolecular interactions/contents in mammalian cell culture harvest.
Abstract:
Histones, in the form of chromatin heteroaggregates in cell culture harvests, are becoming recognized as a major challenge in antibody purification. However, recent publications have shown that immunoassays for host cell proteins do not detect histones. In addition, histone standards provided in commercially available histone ELISA kits are not based on authentic CHO histones. This presentation will show how to prepare authentic host cell histone standards, and how they are necessary for accurate quantitation of histone contamination in cell culture harvests and during purification procedures.
Biography:
Faiz Ali Post Doctoral Fellow advanced separation sciences, graduate school department of chemistry and chemical engineering, Inha University, South Korea.
Abstract:
Miniaturized partially sub-1µm porous silica monolith particles with large pore size, exhibiting excellent chromatographic performance have been synthesized in a relatively large scale by an elaborated sol-gel procedure. The resultant silica monolith particles were chemically modified with chloro-dimethyl-octadecylsilane (C18), and end-capped with a mixture of hexamethyldisilazane and chlorotrimethylsilane. Very good separation efficiency (185,000/m) and resolution (6.05) were achieved when the C18-bound phase was evaluated for a test mixture of five benzene derivatives after packing in a stainless steel column (1.8 mm × 150 mm). The optimized elution conditions were found to be 70/30 (v/v) acetonitrile/water with 0.1% TFA at a flow rate of 25µL/min. The column was also evaluated for fast HPLC analysis using a flow rate of 100µL/min where all the five analytes were eluted in three minutes with still reasonable efficiency (Ca. 96000/m) and excellent resolution within the pressure limit of conventional HPLC. The strategy of particle size reduction combined with C18 modification along with large pores and monolithic architecture has resulted in a useful stationary phase (C18-bound silica monolith particles) of low production cost showing excellent chromatographic performance capable of fast HPLC analysis.
Alberto Chisvert
University of Valencia, Spain
Title: Cosmetic ingredients: From the cosmetic to the human body and the environment
Biography:
Alberto Chisvert (BSc, Faculty of Chemistry, University of Valencia, 1999; PhD, Dept. of Analytical Chemistry, University of Valencia, 2003). He is Associate Professor of the Dept. of Analytical Chemistry of University of Valencia. At present, he has published more than 60 articles in reputed journals, and more than 10 book chapters. Moreover, he is the Co-Editor of the book Analysis of Cosmetic Products (Elsevier, 2007). He is Editorial Board Member of Advances in Analytical Chemistry, American Journal of Analytical Chemistry, Chromatography Research International, International Journal of Analytical Chemistry and Journal of Trace Analysis in Food and Drugs, and is a regular reviewer of more than 27 journals. His research areas are focused on both liquid and gas chromatography coupled to mass-spectrometry, liquid- and solid-phase microextraction, in bioanalysis and cosmetic, pharmaceutical and environmental analysis.
Abstract:
Current social demand for cosmetic products in the developed countries has prompted the cosmetics industry to achieve a leading position within the chemical industry. That is why health authorities are increasingly concerned by this sector, regulating these products, not only to ensure the safety of users but also to ensure their effectiveness. Therefore, quality control in the cosmetic industry is demanded. However, there are not reference analytical methods enough to cover the needs of this sector, so the cosmetics industry is continually requiring the development of analytical methods. Moreover, prohibited substances could be present at trace levels, as they are not intentionally added to the product, but are formed during the manufacturing process. Therefore sensitive analytical methods, in addition to accurate and precise, are needed. Furthermore, studies show that some cosmetic ingredients can be absorbed through the skin and penetrate into the human body, which can be metabolized and/or excreted and even bioaccumulated. Studies are rather scarce in this field as a result of having not analytical methods for determining the cosmetic ingredient and its metabolites at trace levels in biological fluids. Finally, there are several studies that show that cosmetic ingredients reach the aquatic ecosystem causing adverse effects on flora and fauna. Therefore the development of sensitive analytical methods for determining cosmetic ingredients at trace levels on the environment is of great interest from the surveillance environmental point of view. The purpose of this presentation is to provide an overview of the analytical contributions of our group in this three interesting fields.
Katarzyna Owczarek
Gdańsk University of Technology, Poland
Title: Assessment of cytotoxic and endocrine potential of selected xenobiotics commonly present in food products
Biography:
Katarzyna Owczarek has completed her MSc studies in 2014 from Chemical Faculty Gdansk University of Technology. She started PhD studies on Department of Analytical Chemistry in Gdansk University of Technology in 2014. She has published 3 papers in reputed journals and 1 monograph.
Abstract:
More and more increased intensity of anthropo-pressure processes can be observed, among other things, in the release of great quantities of synthetic substances into the environment, including the Endocrine Disrupting Compounds (EDC). A vast number of chemical substances are considered to belong to the this group, including substances which occur naturally in the environment, such as mycotoxins and phytoestrogens, and substances which have been artificially released to the environment as a result of human activities (synthetic hormones, growth promoters, phthalates, bisphenols, metals, pharmaceutical residues, organo-chlorine compounds including PCBs). Taking into account the plurality and chemical diversity of the aforementioned compounds, it should come as no surprise that these substances are almost ubiquitous in various elements of the environment and in the food chain, in which they are subjected to bio-magnification. These chemicals are mainly suspected to contribute to the induction of neoplastic diseases such as the breast and prostate cancer, metabolic diseases, including obesity, genetic modifications and impairment of reproductive functions. They are also suspected to have mutagenic and cytotoxic effects and that they disrupt regulatory pathways of some organs. That is the reason why all attempts to broaden the knowledge of relations and mechanisms triggering the development of relevant diseases are justified. The aim of the conducted research was to evaluate cytotoxicity and endocrine potential of selected EDCs which cause serious food contamination (bisphenol A, 4-nonynphenol, 4-t-octylphenol, diethylstilbestrol, bisphenol A diglycidyl ether and its derivatives and phthalates) with MTT and YES/YAS assays.
Biography:
José Manuel Herrero-Martínez obtained his PhD in Chemistry at the University of Valencia, Spain, in 1996. He worked in teaching and research at the Universities of Amsterdam and Barcelona. Since 2009, he is working as an Associate Professor in the Department of Analytical Chemistry at the University of Valencia. His research interests focus on the development of polymeric materials, in particular polymer monoliths, with application in separation science.
Abstract:
Along the last years, monolithic porous polymers have been widely investigated as stationary phases for separation techniques. Advantages are ease of preparation, high permeability and a vast variety of easily modifiable surface chemistries. Nevertheless, these monoliths have low surface areas due to the lack of an adequate mesoporous structure. Adverse consequences are low retention and reduced sample load capacity. The development of hybrid materials containing nanoparticles (NPs) has opened promising ways of overcoming these limitations, also enormously expanding the available monolith classes. We have work with two approaches, namely to modify the surface of methacrylate monoliths with covalently bonded NPs (I), and to obtain monoliths by copolymerization with derivatized NPs (II). Concerning approach I, the surface of monoliths was first modified with thiol-containing ligands, thus to be able of bonding silver NPs (AgNPs) on them. Monoliths with very large reactive surfaces resulted. The bonded AgNPs showed a large capacity of strongly retain thiol-containing analytes. Further, the retained analytes were easily eluted by replacement with compounds also containing thiol groups if present in excess in the mobile phase. Application to the pre-concentration of thiol-containing compounds (glutathione, N-acetylcysteine, peptides and proteins) in biological fluids and food was demonstrated. Regarding to approach II, iron oxide NPs were first silanized with a vinyl-containing reagent. The resulting vinylized NPs were copolymerized with methacrylate monomers to render a monolith with a rough surface. This monolith exhibited enhanced retention of a variety of molecules (e.g., organophosphorous pesticides), as well as an improved efficiency.
Shobini Jayaraman
Boston University School of Medicine, USA
Title: Comprehensive overview of biophysical studies of lipoprotein stability
Biography:
Shobini Jayaraman has completed her PhD from Indian Institute of Technology, Madras, India and Postdoctoral studies from Weizmann Institute of Science, Israel. She was the recipient of Sir Charles Clore fellowship at Weizmann Institute of Science. Currently she is a Senior Research Scientist at Boston University School of Medicine. She serves as the liaison for academic and industrial contract research services at Boston University. She has published more than 25 papers in reputed journals. Her recent publication in JBC has been chosen as paper of the month in May-2014 by International Atherosclerosis Society.
Abstract:
Lipoproteins are nanoparticles comprised of proteins and lipids that provide vehicles for transport of fat and cholesterol in circulation. High levels of certain lipoproteins increase the risk of heart disease. Each lipoprotein is a non-covalent assembly of several proteins and several hundred lipids. The major challenge in the biophysical analysis of lipoproteins arises from their heterogeneity in size (7-100 nm), density (1.06-1.22 g/L), and protein and lipid composition. Moreover, lipoproteins are highly dynamic assemblies undergoing continuous remodeling via various enzymatic and non-enzymatic reactions. This provides a major challenge for detailed structural studies of lipoproteins. To overcome this challenge, we designed an integrated biophysical approach by combining far- and near-UV circular dichroism (CD) spectroscopy, turbidity, differential scanning calorimetry (DSC), fluorescence spectroscopy, transmission electron microscopy (EM), size-exclusion chromatography (SEC) and other methods to analyze the structure and remodeling of all major lipoprotein classes. This integrated approach was used to study thermal denaturation of human low- and high-density lipoproteins (LDL and HDL, or bad and good cholesterol). The results clearly showed that lipoprotein stability is controlled by kinetics barriers. Interestingly, heat-induced remodeling of all lipoproteins involves partial protein unfolding/dissociation and lipoprotein fusion and rupture. These structural transitions mimic key aspects of in-vivo lipoprotein remodeling. These and other emerging approaches will allow one to study structural, dynamic and functional properties of larger more challenging systems. Ultimately, such integrated approaches are hoped to bridge the gap between the biophysical studies of isolated macromolecules or their complexes, and the complexity of cellular systems.
Yeun-Jun Chung
The Catholic University of Korea, Korea
Title: Stuffer-free multiplex ligation-dependent probe amplification technology for identifying the copy number variation markers associated with the risk of SLE
Biography:
Yeun-Jun Chung is a Professor of Medical Microbiology and Head of Integrated Research Center for Genome Polymorphism (Catholic Medical College, Korea). He has studied genomics based personalized medicine for past twenty years. Since he founded IRCGP, he has studied Variomics and Oncogenomics using array-CGH, GWAS, NGS, MLPA-CE-SSCP and BI tools. He has published >80 research papers including Gastroenterology, Genome Res, Arthritis Rheum, Cancer Res, Bioinformatics, J Pathol etc. Currently he is an editor-in-chief of Genomics & Informatics (official journal of Korean Genome Organization) and an editorial board member of Experimental & Molecular Medicine and World Journal of Gastroenterology.
Abstract:
Copy number variation (CNV) is one of the major components of human genetic variations and it is thought to contribute to inter-individual differences in diverse phenotypes. Several CNVs have been suggested to be associated with systemic lupus erythematosus (SLE) through the target gene approach; however, genome-wide feature of CNVs in the risk of SLE has not well studied. Recently, our group studied the genome-wide CNVs with 964 SLE patients and 711 normal control individuals by whole genome SNP array analysis, and identified three CNVs of RABGAP1L, 10q21.3, and C4 associated with the risk of SLE in Korean Women (Arthritis and Rheumatism 65:1055-63 2013). For the multiplex detection of the 3 CNVs, we adopted a stuffer-free multiplex ligation-dependent probe amplification based on conformation-sensitive capillary electrophoresis, which we developed previously (MLPA-CE-SSCP, Electrophoresis 33, 3052–61 2012). For the MLPA-CE-SSCP based SLE CNV detection system, we also included 3 previously reported CNVs such as CCL3L1, FCGR3B, and EGR2. As a result, all the targets were well separated and the expected CNVs were consistently identified by the SLE MLPA-CE-SSCP system. After validation of the performance of SLE MLPA-CE-SSCP system, we applied the system for 113 SLEs and found that deletion of C4 was the most significantly associated with the risk of SLE. Our results suggest that the SLE MLPA-CE-SSCP system can be useful for predicting the risk of SLE. However, further larger scale study will be required.
Albert Van Den Berg
BIOS/Lab on a Chip group, The Netherlands
Title: Labs, cells and organs on a chip
Biography:
Albert Van Den Berg received his MSc in applied physics in 1983, and his PhD in 1988 both at the University of Twente, the Netherlands. From 1988-1993, he worked in Neuchatel, Switzerland, at the CSEM and the University (IMT) on miniaturized chemical sensors. In 2000 he was appointed as full professor on Miniaturized Systems for (Bio) Chemical Analysis in the faculty of Electrical Engineering. He received several honors and awards such as Simon Stevin (2002), ERC Advanced (2008) and ERC Proof of Concept (2011, 2013) grants, Spinoza prize (2009), Honorary University Professorship (Twente, 2010), and board member of the Royal Dutch Academy of Sciences (KNAW) in 2011.
Abstract:
The recent rapid developments in microfluidic technologies have enabled the realization of miniaturized laboratories. This labs-on-a-chip will play an important role in future medicine, both in point-of-care devices for drug or biomarker monitoring, as well as in early diagnostic devices. We developed a pre-filled ready-to-use capillary electrophoresis platform for measuring ions in blood. It is used to monitor lithium in finger-prick blood of manic-depressive patients, but can also be used for measuring calcium in blood for prevention of milk fever, or for measuring creatinine in blood or sodium in urine for early detection of ESRD. Apart from diagnostic devices, microfluidic devices are increasingly used to realise advance disease and organ-models, as illustrated by the blood-brain barrier chip and a blood vessel on a chip. Finally, a microdroplet platform for encapsulation of single cells in microdroplets, ordering of these microdroplets and 1:1 fusion of these droplets is demonstrated. We believe this is a very powerful new tool that can be used for high-throughput single cell experimentation.
Biography:
Cyril Pernot is working in the compound-semiconductors field since 1997. He received a PhD degree in Physics from the University of Montpellier in 2000 for his work on AlGaN-based UV-photo-detectors. Since 2007 he is involved in the development of AlGaN-based UV-LED and he is currently in charge of the crystal growth section at NIKKISO Co., Ltd for the production of UV LED epitaxial wafers.
Abstract:
Deep UV LEDs (255-350 nm AlGaN-based UV-LED) have many advantages compare to conventional UV lighting such as mercury lamps. As it is a solid-state technology, it has a customizable emission wavelength, long life time, low voltage operation, instant on/off, it is shock resistant, easy to integrate (design flexibility, simple driving circuits), and ecology friendly (no hazardous substance such as mercury, no ozone production). In addition, in recent years performance of Deep UV LED has increased markedly. At NIKKISO, we have overcome many issues plaguing the industry, like output power and lifetime for UVB (280-320 nm) and UVC (<280 nm) LEDs. In this presentation we will review the technical progress that has been made recently, we will present the DUV lighting solutions of Nikkiso (SMD package, Modules), and we will discuss the potential markets penetration including spectroscopy applications in analytical and life sciences instrumentation
Yassine H. Benchekroun
Yassine H. Benchekroun Sidi Mohamed Ben Abdallah University, Morocco
Title: Uncertainty profile; a global strategy for validation and estimation of uncertainty: Application to a high-performance thin-layer chromatographic method
Biography:
Yassine Hameda Benchekroun is a PhD candidate in Chemometrics; His research has focused on several lines: development of methodologies for pharmaceutical and biomedical analysis, and quality assurance for drug assays; chemometric pattern recognition covering methodological developments; experimental design and optimization of the analytical process; validation of analytical methods and estimation of the measurement uncertainty.
Abstract:
The aim of this work is to apply a new technique for the validation of quantitative analytical procedures based on β-content tolerance interval and uncertainty profile. Also, an original strategy for estimating measurement uncertainty by the same approach has been developed. The performance of our proposal was confirmed by application to a High Performance thin-layer chromatography for formulation drugs. Compared to the classical strategy, the new approach has a more holistic character. It means that it is no longer necessary to know the various individual steps into which the analytical method can be broken down since this latter is taken as a whole.
Natalia Szczepańska
Gdańsk University of Technology, Poland
Title: Assessment of toxic effect and endocrine potential of food packages extracts
Biography:
Natalia Szczepańska has completed her MSc studies in 2014 from Chemical Faculty Gdansk University of Technology. She started PhD studies on Department of Analytical Chemistry in Gdansk University of Technology in 2014. She has published 3 papers in reputed journals and 1 monograph.
Abstract:
In the scientific literature related to the widely understood issue of packaging materials designed to have contact with food there is much information on raw materials used for their production, their physiochemical properties, types and parameters. There are also a great number of publications on the management and disposal of used packaging. Unfortunately, not much attention is given to the matters concerning migration of toxic substance from packaging and its actual influence on the body of the final consumer of the food packed, even though health protection and food safety are the priority tasks. The main criterion for approving packaging materials for contact with food are the toxic properties of the substances used to produce a given material and the degree of global migration and specific migration. Unfortunately, however, the standard guidelines for assessing the impact of packaging on food quality only take into account the numerical values of these parameters fixed for a small group of compounds. Such approach raises many objections, given that the results of a recent study showed that apart from the monitored compounds leaching from the surface of the package, there are also other contaminants and their derivatives formed by interaction with food components and as a result of technological procedures applied. The goal of tests was to estimate the impact of foodstuff packaging type, production and storage conditions on the degree of leaching of potentially toxic and endocrine active compounds to foodstuffs with the use of the acute toxicity test Microtox® and endocrine potential YES/YAS.