What's in Your mAb Purification Toolbox?
Mark A. Snyder, PhD
Manager, Process R&D Applications Group
Bio-Rad Laboratories

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Abstract:
As monoclonal antibodies (mAbs) continue to represent a growing portion of development pipelines, downstream process developers are increasingly challenged to design processes that deliver quality product at reduced costs. Because no two antibodies or feedstreams are the same, process developers must continuously expand and adapt their resources to meet the performance goals of their organizations.

Join us for an informative technical webinar to learn about effective tools and new methods that will expand your range of options for mAb purification.

With 25 years of bioprocessing industry experience, a full range (capture to polishing) of media, and multiple FDA approved processes using Bio-Rad products, we are uniquely qualified to help you address your most difficult purification challenges.

Webinar topics include underutilized productivity levers, improved recovery for difficult to elute mAbs, and efficient aggregate and impurity removal.

Antibody Screening and Assay Design
July 23, 2009
Olan Dolezale, PhD
Senior Research Scientist,
CSIRO Molecular and
Health Technologies

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Abstract:
In this free 1 hr webinar, Dr. Olan Dolezal, a Senior Research Scientist at the CSIRO Molecular and Health Technologies, Melbourne, Australia, gives a brief introduction to SPR technology and the ProteOn XPR36 protein interaction array system, followed by an in-depth explanation about how the ProteOn system was used to successfully develop a novel capture assay to be able to screen a library of SvFv fragment antibodies.

To help in troubleshooting, Dr. Dolezal also covers some of his experiences in experiment design. A Q & A session takes place at the end of the webinar.

Epitope Binning Using the ProteOn XPR36 Protein Interaction Array System
June 25, 2009
Yasmina Abdiche, PhD
Senior Principal Scientist,
Bioinnovation and Biotherapeutic
Center, Pfizer Rinat

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Abstract:
In this free 1 hr webinar, Dr Yasmina Abdiche, an expert in the screening and characterization of large molecule therapeutics, discusses how SPR—in particular the multiplex capabilities of the ProteOn system—is used by her lab.

Dr Abdiche describes the development of multiple assay orientations using the ProteOn system that she employs as part of her antibody screening workflow. A brief introduction to SPR and the ProteOn system is included. A Q & A session takes place at the end of the webinar.

An Introduction to the ProteOn XPR36 Protein Interaction Array System
May 27, 2009 Tsafrir Bravman, PhD
Manager,
SPR Applications Group
Bio-Rad Laboratories

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Abstract:
This webinar gives a brief introduction to SPR technology and the ProteOn System and how it compares with other interaction analysis technologies. An overview of how to design an SPR experiment to obtain high quality kinetics follows. Finally, the many applications of the ProteOn XPR36 system, including antibody screening, small molecule-protein interaction, and nucleic acid-protein interaction analysis are discussed.

Proteins — Where DNA Takes on Form and Function
May 15, 2009
Kristi DeCourcy, PhD
Virginia Polytechnic Institute and State University

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Abstract:
Electrophoresis of proteins is similar to electrophoresis of DNA, but there are many differences between protein and DNA molecules including composition, charge, size, and variability that must be addressed to separate proteins by size using electrophoresis. This webinar will cover the similarities and differences between DNA molecules and protein molecules and the changes in procedure that are necessary to deal with these differences. After size separation, proteins must be visualized with a dye and the result analyzed, but what information other than size can be gained from a protein gel? Further experiments such as western blotting will be discussed.

About the speaker:
Dr. Kristi DeCourcy received her BA in biology from SUNY Purchase and her MS in biology and PhD in biochemistry from Virginia Tech. Her doctoral research focused on protein chemistry and cell biology. For the last 14 years, she has been the laboratory manager of the Fralin Biotechnology Center at Virginia Tech. As manager, Dr. DeCourcy has run the undergraduate molecular biology laboratories, managed a microscopy core facility, and developed and directed a high school outreach program that provides equipment and materials to more than 10,000 Virginia students annually.

From Plants to Sequences: A Six-Week College Biology Lab Course
May 16, 2008
Dr. David Robinson
Dr. Joann Lau

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Abstract:
In this FREE web conference, we will share the Cloning and Sequencing Explorer Series for student research developed in collaboration with Bio-Rad Laboratories (available Fall 2008).

Gene-cloning techniques are a common topic in most molecular biology courses, but providing students with hands-on experience is more challenging. The objective of this project is to have students isolate, sequence and characterize a pivotal gene of glycolysis (Glyceraldehyde 3-Phosphate Dehydrogenase; GAPDH) from a plant species, or cultivar, not previously published. In addition to wet-lab techniques (e.g. DNA extraction, PCR, ligation, transformation, restriction digest), students gain experience with bioinformatics analysis of their unique clones. This includes BLAST, identification of introns/exons, contig construction, phylogenetics, and acquaintance with different bioinformatics databases. These results are publishable in the NCBI GenBank.

About the speakers:
Dr. Dave Lowell Robinson received his B.S. and M.S. degrees in Plant Sciences from the University of Arizona, and his Ph.D. in Plant Physiology from the University of Minnesota. He has been teaching subjects like Botany and Molecular Biology at Bellarmine University (Louisville, KY) for the past 13 years. He has published 18 refereed journal articles on topics in cellular/molecular biology, weedy-plant physiology, and biological education, and has published 13 DNA accessions in the NCBI GenBank with student co-authors. Several educational exercises have been developed in his lab as a result of his role as Principal Investigator on a NIH grant meant to enhance biomedical education within the state of Kentucky.

Dr. Joann M. Lau received her B.A from Bellarmine University, and recently her Ph.D. from the University of Illinois at Urbana-Champaign. Her graduate work focused on developing plant based edible vaccines for the Respiratory Syncytial Virus (RSV). During her graduate studies at Illinois, she was the recipient of the Department's Eugene S. Boerner Graduate Fellowship, the Agricultural Genome Sciences and Public Policy Graduate Traineeship sponsored by the USDA, the Colgate-Palmolive Fellowship, as well as the Society for In Vitro Biology Student Award. Joann teaches Drugs and the Human Body, Molecular Biology, and a variety of laboratory courses at Bellarmine University. She has mentored numerous successful undergraduate research projects on topics as diverse as biomedical genetics, phylogenetics, bioinformatics, plant transformation, and the use of plant derived-products for the treatment of cancer. In the future, she would like to continue developing curricular materials using the latest biological techniques.

Tackling Reproducibility Issues in Mass Spectrometry-Based Biomarker Discovery
December 5, 2007
Dr. Martin Latterich
Dr. Timothy D. Veenstra
Dr. Toni Whistler

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Abstract:
Mass spectrometry is an extremely powerful and sensitive technology that can detect very small changes in protein expression levels. Some of these changes may stem from the biological differences related to a disease or treatment of interest. Others, however, may reflect the heterogeneity of patients across multiple sites, the inherent biological complexity and diversity of different sample types, and even small differences in sample collection, processing, handling, and analysis techniques used by multiple operators across multiple locations. As a consequence, data may be tainted by site-, study-, population-, or sample-specific anomalies and, therefore, not be sufficiently robust for biomarker discovery.

About the speakers:
Dr. Martin Latterich has over 14 years' experience in pharmacogenomics, proteomics, and the biochemistry of proteins and small molecules. Dr. Latterich currently holds a faculty position jointly at the Montreal Heart Institute and the University of Montreal, Faculty of Pharmacy. He previously served on the faculty of McGill University in Montreal as a Tier 1 Canada Research Chair and the Salk Institute in San Diego, and he has made significant contributions to the field of integrative proteomics and pharmacogenomics through the use of innovative mass spectrometry and biochemical techniques. His grant-funded work has been recognized by the 1998 Pew Scholar Award and the 1997 Basil O'Connor Starter Scholar Award.

Dr. Timothy D. Veenstra is Director of the Laboratory of Proteomics and Analytical Technologies (LPAT), SAIC-Frederick, Inc. at the National Cancer Institute at Frederick, MD. Dr. Veenstra joined NCI-Frederick in the fall of 2001 to implement a state-of-the-art mass spectrometry center that focuses on the discovery of signaling pathways and networks that are activated within cells as a response to external perturbations. His laboratory is also keenly interested in utilizing high-throughput proteomic approaches for the discovery of biomarkers for diseases such as cancer and exploring the use of proteomic and metabolomic technology for the routine interrogation of clinical samples.

Dr. Toni Whistler is a Senior Service Fellow in the Chronic Viral Diseases Branch, Division of Viral and Rickettsial Diseases, at the Centers for Disease Control and Prevention. Dr. Whistler has extensive experience in setting up mass spectrometry analysis, particularly SELDI-TOF. She has a strong publication record, including two book chapters and recent papers on the optimization of SELDI-TOF mass spectrometry experiments. Dr. Whistler has received a number of awards for her research, notably the HHS Secretary's Award for Distinguished Service from the Department of Health and Human Services in 2000.

Using Multiple Reference Genes and Other Factors to Obtain High-Quality Gene Expression Data
August 27, 2007
Dr. Jo Vandesompele

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Abstract:
Real-time PCR is the current standard for accurate, sensitive, and fast gene expression measurement. The reliability of the final quantitative results is heavily dependent on the quality of all elements in the laboratory and data analysis workflow, including the input template (integrity and absence of inhibitors), the PCR assay (specificity, efficiency, limit of detection), experimental design, normalization strategy (use of multiple validated reference genes) and data processing steps. In this webinar, Dr. Vandesompele illustrates these important issues, with a specific focus on the identification, use, and utility of multiple reference genes to obtain high-quality gene expression results.

About the speaker:
Dr. Jo Vandesompele earned his Master's degree in bioengineering (MS BioE) in 1997 from Ghent University, followed by a PhD in medical genetics in 2002. Currently, he is a postdoctoral fellow at the Center for Medical Genetics in Ghent, Belgium. He has coauthored over 60 papers in international peer-reviewed journals. The main themes of his research program are the establishment of genetic and biological bases of childhood tumor neuroblastoma, methodological and technical developments in the field of real-time quantitative PCR, and development of bioinformatic tools such as geNorm, qBase, RTPrimerDB, arrayCGHbase, methBLAST, methPrimerDB, PGE, and Neuroblastoma Gene Server.

Experimental Strategies for Biomarker Discovery and Development
August 22, 2007
Dr. Diane McCarthy

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Abstract:
Typical biomarker studies can be divided into four phases: discovery, validation, purification and identification, and assay development. We will discuss these phases and present approaches for optimizing biomarker workflows to deliver robust biomarkers and biomarker assays. Case studies using the ProteinChip SELDI system in both preclinical and clinical areas will be presented to illustrate optimized biomarker workflows.

In the discovery and validation phases, it is especially important to optimize study design and statistical methods to avoid preanalytical bias and to yield robust markers. Identification can be performed at the end of the discovery phase or at any point during the validation phase. Identification generally requires standard protein purification procedures, such as column chromatography, size filtration, and SDS-PAGE, followed by protease digestion and sequence analysis on a tandem mass spectrometer. Identification of biomarkers provides insight into disease biology and facilitates the development of analyte-specific assays. Once the biomarkers have been positively identified, quantitative assays can be developed for routine testing on an appropriate immunoassay platform (SELDI, Bio-Plex suspension array system, or ELISA). SELDI-based immunoassays are particularly useful for detecting biomarkers with posttranslational modifications.

About the speaker:
Dr. Diane McCarthy oversees the operation of Bio-Rad's Biomarker Research Centers. She received her PhD in biochemistry from The University of Texas at Austin, and she completed a postdoctoral fellowship focused on protein expression, purification, and structural analysis. She has over 5 years of experience managing biomarker research collaborations with leading academic groups (including the National Cancer Institute and Johns Hopkins University) and pharmaceutical companies (including publicly disclosed collaborations with Novartis, Bayer, and Sanofi-Aventis).

From Singleplex to Multiplex: Making the Most Out of Your Real-Time Experiments (Presented in Mandarin)
August 10, 2007
Dr. Feng Pan

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Abstract:
Real-time PCR, a powerful advancement of the PCR technique, quantitates the initial amount of a template with more specificity, sensitivity, and reproducibility than any other method. It has been widely used for gene expression, genotyping, pathogen detection, and many other applications. In this webinar, Dr. Pan presents ways to achieve optimal sensitivity and reproducibility with every assay, from single-target amplification using SYBR Green I to multiple-target amplification (multiplexing). Every laboratory can benefit by using these methods to achieve better results.

About the speaker:
Dr. Feng Pan has been supporting real-time PCR applications for two years as a field application specialist at Bio-Rad Laboratories. Before that, he worked as a postdoctoral fellow in the department of cell biology at Harvard Medical School. He earned his PhD in biochemistry from the University of Rhode Island. Dr. Pan grew up in China and went to Nankai University, where he earned his BS in genetics.

Affinity-Tagged Protein Purification With an Automated System Increases Access to Purified Recombinant Proteins
May 23, 2007
Dr. Laurie Usinger

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Abstract:
Affinity-based protein purification is an increasingly common method used for generating milligram quantities of target protein for use in functional and structural experiments. The Profinia protein purification system has been developed to automate affinity purification and desalting of tagged proteins. The system requires minimal user interaction and can purify target protein in quantities of 1–100 mg in 30 minutes. The instrument uses optimized, preprogrammed methods and prefilled reagents and cartridges to provide fast, reproducible, and cost-effective protein purification. In this presentation, examples of both histidine- and GST-tagged protein purification will be shown to demonstrate the consistent results that can be obtained from using automated purification methods. In addition, hands-on labor time will be compared with traditional techniques, such as gravity flow columns. Improvements in the speed and automation of routine affinity purification can improve laboratory efficiency and the consistency of results across multiple users. The Profinia system increases the ease and reproducibility of affinity techniques so all members of a laboratory can produce quality target protein.

About the speaker:
Laurie Usinger, PhD, was the lead scientist for the Biotechnology Explorer program at Bio-Rad Laboratories from 1996 to 2001, and was instrumental in the launch of seven classroom kits in the first year of the program. The kits teach the basic workflows of molecular biology, from gene expression to protein purification, and use Green Fluorescent Protein (GFP), expressed from the pGLO plasmid, as the primary teaching tool. Dr. Usinger has since held the following positions at Bio-Rad:

• Lead scientist — Precision Protein standards project (under the guidance and management of Roger Provost), 1998–2000
• Project manager and lead scientist — Development and launch of Precision Plus Protein standards project, 2002
• Project manager — Launch of Precision Plus Kaleidoscope standards, 2004
• Program manager — Profinia protein purification system, 2004–2006
• Lead scientist — Construction and design of consumables line supporting the Profinia protein purification system and management and coordination of the consumables with the instrument platform, 2004–2006

Multiplex Innate Cytokine Signal Analyses of Brain Function
April 26, 2007
Dr. Richard P. Kraig

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Abstract:
Cytokines are powerful signaling molecules that are increasingly recognized for their involvement in normal brain function. Our goal is to decipher how innate cytokine changes associated with increased neural activity, such as from learning, make the brain more resilient to disease. This pursuit is complicated by the complex interactive behavior of innate cytokine signaling, which is highly redundant and pleiotropic — multiple cytokines can have the same effect, and a single cytokine can have variable effects. Our approach is to simultaneously measure key signaling protein changes via multiplex bead-based assays using the Bio-Plex suspension array system. We use these data to define critical node signaling sites of the innate cytokine signaling network involved in activity-dependent neuroprotection. The application of this experimental strategy to the brain, which is regionally and cellularly heterogeneous, will be discussed.

About the speaker:
Dr. Richard Kraig, MD, PhD is an authority on the role of glial cells in the brain, the treatment of migraine headaches, and how the brain protects itself from injury. An active researcher, educator, and clinician, Dr. Kraig is the William D. Mabie Professor in Neurosciences at the University of Chicago Medical Center. His laboratory research is focused on examining mechanisms by which the brain naturally protects itself, and how physical, mental, and social activities can lessen the impact of neurological disease.

Dr. Kraig earned his BA in chemistry from Cornell College, his PhD in physiology and biophysics from the University of Iowa, and his MD from New York University. He was an intern at the University of Chicago and did his residency in neurology at the New York Hospital-Cornell Medical Center, where he took his first faculty position before joining the University of Chicago faculty in 1988.

Moving xMAP Technology Forward: New Targets and High-Sensitivity Precision Pro Assays
March 30, 2007
Dr. Deyrick Dean

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Abstract:
Increasing interest in using xMAP technology as an alternative to ELISA and western blot approaches has fostered a demand for new assays that are suitable for a broad range of research applications. This need is not simply limited to new targets. As xMAP technology is more widely adopted, there is an added requirement for assays with higher performance and automation capability. This presentation will discuss the use of the Bio-Plex suspension array system, an integrated open platform for xMAP technology. An overview of the system and its unique set of software tools for instrument control, analysis, and validation, as well as assay options, will be discussed. Focus will be given to applications of xMAP technology and data generated using the Bio-Plex system. In addition, results using a specialized panel designed to address the xMAP user's needs for higher sensitivity and precision when measuring cytokine levels will be presented.

About the speaker:
Dr. Deyrick Dean began working with xMAP technology over six years ago while conducting research at a biotech start-up company in Winston-Salem, North Carolina. During the past four years with Bio-Rad Laboratories, he has continued to expand his base of knowledge. In his current role as Bio-Plex program manager, USA, he frequently travels to present technical seminars, consult with researchers, and train laboratories using xMAP technology.

Exploring One-Shot Kinetics and Small Molecule Analysis Using the ProteOn XPR36 Protein Interaction Array System
November 15, 2006
Dr. David Myszka

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Abstract:
A ProteOn XPR36 protein interaction array system was used to characterize a set of small molecule–enzyme interactions. With a single injection into the ProteOn system's crisscross flow system, response data was collected for six concentrations of each analyte over six target protein surfaces. This "one-shot" approach to kinetic analysis improves throughput while generating high-quality data. Results illustrate the potential of this parallel-processing biosensor to increase the speed of kinetic analysis in drug discovery and to expand the applications of real-time protein interaction arrays.

About the speaker:
Dr. David Myszka is the founder and director of the Center for Biomolecular Interaction Analysis at the University of Utah.