Genetic Engineering & Biotechnology News

SEP15 2017

Genetic Engineering & Biotechnology News (GEN) is the world's most widely read biotech publication. It provides the R&D community with critical information on the tools, technologies, and trends that drive the biotech industry.

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Genetic Engineering & Biotechnology News | | SEPTEMBER 15, 2017 | 11 • The matrix effect: Unlike singleplex assays, the accuracy of multiplex systems may be adversely affected by the mixture of multiple antibodies and calibration standards. • Reagent lot-to-lot variability and kit reproducibility. Dr. Chimento: In the multiplex assay, the re- agents must work together correctly on the same sample matrix, and they must function on the desired platform. A common approach in developing a multiplex assay is to purchase or license commercially available reagents. The challenge comes because the reagents were not specifically designed to function to- gether or specifically on the sample type, or for the platform being used by the researchers. The outcome is a multiplex with specific gaps for the missing reagents. The researchers may then need to develop "fit for purpose" reagents to fill the gaps in the multiplex assay. GEN: Have there been recent scientific discoveries and advances that made a significant impact on the develop- ment of multiplex immunoassays? Dr. Pregibon: To meet the diverse needs of drug discovery and biomarker development, immu- noassays have seen advances in sensitivity for single proteins, assay simplification for high- throughput automation, and the use of both immunoassays and nucleic acid assays, as well as spatial profiling. Platforms capable of profil- ing both proteins and nucleic acids, including NanoString's Vantage, Bio-Techne's Luminex, and Abcam's FirePlex, are finding increased use in research and clinical laboratories. Dr. Tan: The key sectors that have made a significant impact on the development of multiplex immunoassays are data science, biology, and instrumentation. The data sci- ence or bioinformatics sector is driven by the demand for big data integration with expert knowledge–driven approaches to develop biologically relevant assays and to better as- sess the complexity of multifactorial diseases such as cancer, stroke, and diabetes mellitus. The biology sector is fueled by biomarker discovery, drug development, and the emerg- ing demands for multiplex point-of-care test- ing. The instrumentation sector is driven by the demand for automation and improvement in detection capability of low-abundance targets. Ms. Johnson: The advances made over the past decade are allowing science to move at a much faster pace. Our customers' priori- ties and projects can change quickly, so we need to be agile in our ability to deliver the multiplex assays they need. This has led to a shift toward a developmental pipeline that is more responsive (customer request) than preplanned (literature research). We often have customers that are happy with our Luminex ® Performance Assays but want additional analytes, customized assay specifications, etc. Our assay development expertise and reagent inventory have al- lowed us to respond to customer needs. Dr. Wild: The development of multiplex as- says that can be run on standard equipment such as flow cytometers expands the range of applications of multiplex assays. Efforts in improving antibody quality and standard- ized production improves the reliability and comparability of results. In addition, we ex- pect multiplex assays to speed up discoveries in new research topics such as extracellular vesicles (exosomes). Dr. Mao: Currently, less than 10% of the hu- man proteome can be measured in a quan- titative manner. One of the limiting factors is the availability of high-quality antibodies. To overcome this bottleneck, small synthetic molecules that have antibody-like function- ality are garnering more attention, including DNA aptamers, antibody mimics, and pro- tein scaffolds. Antibody colocalization microarray tech- nology, wherein individual detection antibod- ies are printed directly atop their correspond- ing capture antibodies, has been explored as a solution for cross-reactivity among an- tibodies. Finally, DNA amplification–based proximity ligation (PLA) technology has been employed to increase immunoassay de- tection sensitivity. Dr. Chimento: Technological developments within both ancillary reagents and plat- forms have made a major impact on multi- plex assays. Robust antibody reagents have been around for some time. Now they may be combined with (1) highly specific fluo- rochrome or dye-based reporters (which may be used with antibodies and beads), (2) exceptional microfluidics, and (3) out- standing detection sensors. These reporter and detection system combinations have made it possible to create not only a multi- plex assay but even a single-well multiplex assay, where multiple reporters are deliver- ing data simultaneously, reducing costs and saving time. Drug Discovery C L A R I O s t a r ® w i t h A C U Full fl exibility in cellular assays with the new gas ramping function For the fi rst time scientists are given the capability to fully manipulate the environment within the microplate reader, by mimicking in vitro hypoxia, ischaemia/reperfusion and much more. The CLARIOstar with LVF Monochromators TM is not only the most- sensitive monochromator-based plate reader today. Its Atmospheric Control Unit (ACU) can run gas ramps, modifying O 2 and CO 2 gas levels and rapidly restoring back atmospheric conditions. 0 20 40 60 80 100 120 Time in minutes Gas concentration in % 20 18 16 14 12 10 8 6 4 2 0 CO 2 O 2 Ischaemia Reperfusion Example of O 2 deprivation and reoxygenation (down to 0.2% O 2 ; purple) with steady 5% CO 2 (grey) performed by the CLARIOstar with ACU. © 2017 All rights reserved. All logos and trademarks are the property of BMG LABTECH.

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