Genetic Engineering & Biotechnology News

AUG 2017

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Genetic Engineering & Biotechnology News | GENengnews.com | AUGUST 2017 | 13 Microplate Reader and Imager Software read- ily identifies each object to generate HC cell counts that are comparable to counts achieved using nuclear labels, across a wide range of cell densities (Figure 1). Combining this pow- erful label-free cell counting technique with the temperature, humidity, and CO 2 controls available on the Lionheart FX and Cytation systems enables uncompromising long-term kinetic proliferation studies. Additionally, coupling the Cytation with the BioSpa 8 Au- tomated Incubator provides a fully automated system for conducting proliferation studies on up to eight cell culture plates at a time. Kinetic Label-Free Proliferation Assay The HC Cell Counting kinetic prolifera- tion assay enables quantitative evaluation of cell-division rates for normal and malignant cell types, as well as measuring effects of antiproliferative treatments (Figure 2). For many cell types, a proliferation assay lasting three to five days is sufficient for capturing multiple population doublings and gener- ating meaningful data. Initial cell densities should take into account cell-doubling time and duration of experiment. A four-day experiment with 2,000 cells seeded per well in a 96-well microplate is a good starting point for cell lines with a dou- bling time of around 24 hours. These param- eters result in robust counts while establish- ing a low enough cell density that the culture will not become fully confluent until well into the experiment. Environmental condi- tions, including temperature (37 o C), gas (5% CO 2 ), and humidity (90%) are maintained throughout the assay. During the automated protocol, two im- ages are collected for each well every two hours—one high-contrast brightfield image for reference and qualitative evaluation, and one HC Cell Count image. Gen5 image pro- cessing and analysis is done in real time while the assay is running. Although image-analysis settings have been optimized for a variety of cell types, all settings should be determined empirically by visually confirming accurate placement of object masks using a few rep- resentative images. If modifications to the im- age-analysis settings are required, adjustments to the threshold value and object size should be sufficient to address most conditional variations. Once settings are confirmed, the protocol can be saved for future use with the same or similar cell types. Unique Biological Insight into Drug Treatment Effects Calculating IC 50 values are a useful met- ric for quantifying dose response. However, values based on a single endpoint can yield incomplete and misleading results. Kinetic proliferation assays provide a detailed pro- file of the effect of drug treatment, enabling researchers to evaluate treatment response over time and determine the optimal time course for analysis. Using the area under the curve (AUC) from each proliferation profile accounts for dynamic drug-treatment effects, resulting in a more relevant evaluation of dose response and drug kinetics (Figure 3A). An additional advantage of this imaging- based kinetic assay is the detailed visual re- cord of cell health and morphology that is available for deeper analysis. This informa- tion can provide valuable mechanistic insight into cellular responses and reveal more sub- tle drug-treatment effects (Figure 3B). Conclusions HC Cell Counting provides an accurate and convenient method to quantify the size of a cell population without the need for fluorescent labels. Using this technique to conduct kinetic cell proliferation as- says on the Lionheart FX and Cytation BioSpa system enables long-term, detailed evaluation of cell-population viability and growth. View It Now! On Demand DURATION 60 minutes COST Complimentary Speakers Win More Biologics Development Battles with Early Stability Analysis Biophysical and chemical properties are critical factors that can dictate success or failure in the development of therapeutic antibodies. These attributes and all the functional characteristics of antibodies depend on amino acid sequence, post-translational modifications, and the solution environment. Early quality control and risk assessment of biophysical parameters help prevent failure in later stages of antibody development. An early-stage developability workflow was designed to set stage gate for molecules progressing to late-stage development. Application of the Uncle platform in antibody selection will be presented in this webinar. Stability analysis of engineered antibody variants was evaluated using Intrinsic Fluorescence Conformational Stability (T m ) and Colloidal Stability (T agg ). B 22 /k D parameters for monoclonal antibodies (mAbs) in solution were determined and validated by Small-Angle-X ray-scattering (SAXS). Quantitative correlation between T agg and particle size for mAbs was observed in this study. A live Q&A session will follow the presentations, offering you a chance to pose questions to our expert panelists. Who Should Attend • Researchers developing monoclonal antibodies • Drug discovery scientists • Investigators interested in biologic stability • Biochemists • QC/QA scientists Free Registration! www.GENengnews.com/stability-analysis Webinars You Will Learn • How to use T m and T agg measurements to evaluate the stability of engineered antibodies • How the aggregation propensity parameters B 22 and k D can help further evaluate antibody variants • How the Uncle platform can collect multiple measurements earlier, with less sample Dina Finan, Ph.D. Product Manager Unchained Labs Sam Wu, Ph.D. Principal Scientist Janssen BioTherapeutics Produced with support from Drug Discovery Tutorial Joe Clayton, Ph.D. (customercare@biotek. com), is principal scientist at BioTek Instruments. Website: www.biotek.com.

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