Innovative Approaches to Investigating KRAS Therapeutics with Biochemical and Cell-Based Assays
(June 15, 2022)
Kirsten rat sarcoma viral oncogene homologue (KRAS) is the most frequently mutated oncogene, present in approximately 25% of tumors. Numerous studies on KRAS in the initiation and advancement of cancer are constantly updating our knowledge of the gene. Despite drug-targeting attempts over the past four decades, KRAS has been labeled a difficult therapeutic target, even ""undruggable”, however, recent advances, such as the FDA approval of Amgen’s sotorasib, exemplifies and encourages new progress in the field.
This webinar explores innovative methods to discover and qualify KRAS inhibitors with downstream signaling and binding assays, utilizing no-wash HTRF® and AlphaLISA® technology to further the discovery of drugs targeting KRAS.
What You Will Learn:
• How to screen compounds with binding to identify positive hits and competitors
• Detection of MAPK activation to evaluate the efficacy and potency of KRAS inhibitors
• Data supporting indirect targeting of KRAS inhibitors with the RAF MEK approach and direct targeting of KRAS SOSI and KRAS G12 inhibitors
Macrophages: Hideouts for HIV and Drivers of Inflammation
(May 31, 2022)
Human immunodeficiency virus (HIV) is a chronic infection that causes acquired immunodeficiency syndrome (AIDS) and currently infects over 38 million people. Despite the success of antiretroviral therapy (ARV) in controlling HIV infection, almost 700,000 people died from AIDS-related deaths in 2019. For people who have access to ARVs (only 66% of people living with HIV), infection-associated inflammation persists, increasing the risk of developing co-morbidities such as cardiovascular disease and HIV-associated neurocognitive disorder. This emphasizes the need to study the pathogenesis of HIV to aid the development of cure strategies.
Unlike viruses that cause acute infections, like influenza or SARS-CoV-2, the immune system is unable to eliminate the virus from the body. Understanding what cells are infected by the virus and how killer immune cells interact with them will help us understand why HIV persists. While white blood cells called CD4+ helper T cells are the main cell type infected by HIV, tissue-resident phagocytes, called macrophages, also become infected and persist despite ARV treatment. Our lab is studying how macrophages persist in the presence of killer immune cells.
Key Learnings:
● White blood cells that can kill cancer and virus-infected cells, including CD8+ T cells and natural killer (NK) cells, experience more difficulty with killing HIV-infected macrophages compared to HIV-infected CD4+ T cells.
● The poor killing of HIV-infected macrophages further drives inflammation that could contribute to the development of co-morbidities.
● Current efforts are focused on studying the mechanisms of how macrophages inhibit CD8+ T cell and NK cell killer activity.
How to eliminate false-positive ELISA results in clinical samples
(May 23, 2022)
Learn to tackle interference from rheumatoid factor and heterophilic antibodies to achieve trustworthy ELISA results.
The webinar kicks off with a presentation by Dr. Natalie Sippl, Ph.D. in rheumatology, followed by a panel discussion.
The Ultimate Biophysics Tool: Fast and Accurate In-solution Analysis of Proteins and Nanoparticles
(Apr 27, 2022)Fida 1 is a versatile, easy-to-use platform to
give you fundamental, quantitative information about the samples you are working with, including size, polydispersity, environmental stability, aggregation, viscosity, labeling efficiency, Protein Database Bank correlation, etc. It only takes 4 minutes, and you only need four μL of sample material. The samples are analyzed “as is”, i.e., labeled or label-free, no immobilization, no buffer constraints, temperatures from 4-55 deg. C, purified or non-purified, etc.
Supported by dedicated software, Fida 1 also offers access to advanced analyses of, amongst others, membrane proteins, targeted protein degradation, liquid-liquid phase separation, VHH clone selection, bi-specifics, immunogenicity, exosomes, AAVs, etc.
Though a new technology, the Fida 1 is already adopted by several large pharmaceutical companies and leading international research institutions.
Key Learnings:
• Flow-induced dispersion analysis is a unique way of getting fundamental information about parameters essential for understanding the entities you are working with.
• Fida 1 enables advanced analysis of complex constructs and complicated interactions.
• The use of the Fida 1 platform is supported by easy-to-use software. To generate data requires only a few hours of training.
AI-Designed Mutation-Resistant Broad Neutralization Antibodies Against Multiple SARS-CoV-2 Strains
(Apr 6, 2022)
Artificial intelligence has the advantage of allowing for the
analysis of big and ever-changing data such as the dynamic genome information of a constantly evolving virus. In this study, we have applied AI technologies in the computational design of broad neutralization antibodies against over 1300 different historical SARS-CoV-2 strains with very small computational cost. The AI-designed antibodies were tested in vitro and demonstrated high potency against multiple strains in pseudo-virus and real virus neutralization assays. These AI-designed antibodies also exhibited a very high cross-binding hit rate against different RBD mutants in ELISA assays. The results shed light on future therapeutic designs for pandemic preparedness. The study also indicates that there are hidden patterns in viral evolution and that these patterns can be learned by AI to design antibodies against current and future mutant strains within certain evolutionary windows.
Key Findings:
• AI can allow for the design antibodies with in vitro efficacy and drastically reduce the time and cost of antibody engineering such as affinity maturation.
• AI can allow for the design of cross-binding antibodies against a large number of different antigen population such as viral mutant strains.
• AI can reveal patterns of viral evolution processes and allow for the design of antibodies against future viruses beyond current strains.
Structure-Based Charge Calculations for Predicting Properties and Profiling Antibody Therapeutics
( Mar 16, 2022 )
Sino Biological present a method for modeling antibodies and performing pH-dependent conformational sampling, which can enhance property calculations. Structure-based charge descriptors are evaluated for their predictive performance on recently published antibody pI, viscosity, and clearance data. From this, we devised four rules for therapeutic antibody profiling which address developability issues arising from hydrophobicity and charged-based solution behavior, PK, and the ability to enrich for those that are approved by the U.S. Food and Drug Administration. Differences in strategy for optimizing the solution behavior of human IgG1 antibodies versus the IgG2 and IgG4 isotypes and the impact of pH alterations in formulation are discussed.
Key learnings:
How Octet® BLI works in therapeutic discovery and development, including common challenges and solutions
Adoption of the Octet® BLI in the development of advanced therapeutics such as Bispecific antibodies, Nanobodies and Antibody-drug conjugates
Benefits of using a real-time label free analytical tool such as the Octet® BLI relative to ELISA and SPR
Best practices and protocols to generate high-quality data quickly and easily using the Octet® BLI platform
Key Learnings:
• Methods for generating 3D Antibody Models from 2D protein sequence information
• Computational approaches for assessing the developability of Biotherapeutics
• Application of protein descriptors for biophysical property predictions
• Rules for profiling antibody therapeutics
• Considerations for subclass and isotypes in solution behavior and formulation
Bridging In Vitro Data to Clinical Outcomes with MetID
(MAR 8, 2022)
Profiling, characterising, and identifying metabolites are key steps in supporting drug discovery and development of novel compounds. Metabolite identification (MetID) studies help you to understand presence of potential active or reactive metabolites and observe the unique metabolites in your compound to better understand your molecule’s biotransformation capabilities. SEKISUI XenoTech and Agilex Biolabs’ combination of in-depth scientific knowledge in drug metabolism and mass spectrometry, extensive pharmaceutical industry experiences, and advanced software, can quickly identify metabolites in both in vivo and in vitro samples and help accelerate sponsors’ drug development programs through FIH clinical trials.
Key discussions:
• How to improve bioanalytical clinical outcomes by performing MetID characterisation.
• How metabolite characterisation helps troubleshoot problems earlier in the drug program.
• Test system selection for in vitro MetID.
• The use of MetID for tox species selection.
• Integrating MetID with your bioanalytical program and how the two corelate.
• Why metabolite characterization studies are not quantitative.
• Regulatory expectations for MetID.
Reactive Metabolite Detection Studies: Cysteine Trapping by Xenotech
(JANUARY 27, 2022)
About the webinar: Drug-induced liver injury (DILI) is caused by various mechanisms, so conducting a variety of experiments to form a comprehensive score leads to more accurate risk assessment. Including covalent binding for reactive metabolite detection in this approach greatly improves selectivity and specificity, however, that requires a radioisotope-labeled test article, making it unsuitable for early stage development. Glutathione (GSH) trapping is often conducted in its place, but that can be costly with lower throughput or less quantitativity, depending on the trapping agent.
The cysteine trapping study developed by our sister SEKISUI lab, the Drug Development Solutions Center (DDSC), is more quantitative with higher throughput and lower cost. Listen to a Xenotech expert discuss:
* The relation between DILI and reactive metabolites
* Evaluating reactive metabolites
* A relevant case study
* A comparison of different GSH and cysteine trapping evaluation methods
* Their cysteine trapping study design and assay procedure
* Comparative data
About the presenter:
Miki Fujishima is an Assistant Manager in the Business Development department at the Drug Development Solutions Center in Tokai, Japan. In this role, she facilitates ADME solutions for clients and works closely with research & development scientists to develop and improve assays relating to liver toxicity. Miki received her master’s degree in Drug Metabolism and Molecular Toxicology from Tokyo University of Pharmacy and Life Sciences in 2007. She joined the Drug Development Solutions Center in 2007, helping to establish Drug Transporter study offerings performed at both the Tokai facility and SEKISUI XenoTech. She moved on to specialize in DMPK test systems, including hepatocytes and subcellular fractions, and finally to her current position assisting clients in their assessment of Hepatotoxicity.
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Ultra-Efficient Pharmacokinetic Measurements in an Automated, Microscale Format
(JANUARY 26, 2022)
About the webinar:
Pharmacokinetic and pharmacodynamic studies form an important foundation for drug development by profiling therapeutic agents and their effects across multiple timepoints. Unfortunately, PK/PD protein measurements are often limited by plate or bead-based methods with narrow dynamic range, relatively large volume requirements, and long assay times. These factors restrict both the quantity and the quality of proteomic data that can be collected from cell-based or animal models. Correlia offers an automated platform to overcome these bottlenecks - enabling highly efficient protein measurements from as little as 2 microliters of a diluted biosample. In this webinar, we focus on case studies in therapeutic antibody development. Both general and specific measurements of monoclonal antibodies are made possible, respectively based upon IgG-generic and ligand-specific binding affinities.
Key Takeaways
• Protein concentrations can be measured in a fraction of the time and sample volume required for conventional immunoassays.
• Targets are measured in a “sample-in, answer-out” workflow with minimal user intervention.
• Correlia measurements demonstrate extended dynamic range when validated against competing immunoassays.
About the speaker:
Samuel Tia is a co-founder and CSO of Correlia Biosystems, where he works with their scientific teams to expand Correlia’s immunoassay test menus and deliver solutions for customer proteomic applications. Dr. Tia received a Ph.D. in Bioengineering from UC Berkeley, where his work in the Herr Lab led to a licensed patent. Prior to graduate studies, he obtained a BS in Mechanical Engineering from the University of Florida and had industry experience in drug delivery and medical-grade manufacturing/packaging.
Bead Assisted Mass Spectrometry (BAMS): A High-Resolution, Immunoassay Platform for Multiplexed Biomarker Screening
(Oct 13, 2021 02:00 PM in Eastern Time, US and Canada)
The ability to quantitatively probe diverse panels of proteins and their post-translational modifications (PTMs) across multiple samples would aid a broad range of biological, biochemical, and pharmacological research efforts. A novel microarray-based immunoassay technology that can support highly multiplexed, targeted proteomic studies has been developed by Adeptrix. The technology, termed Affinity-Bead Assisted Mass Spectrometry (Affi-BAMS), combines immunoaffinity capture with mass spectrometry detection, and it enables the development of highly specific and customizable assay panels for simultaneous profiling of multiple proteins, PTMs, and proteoforms. The Affi-BAMS workflow uses a bead-based enrichment approach in which each bead contains a single, protein (or site) specific antibody. Multiplexing capability is achieved by combining Affi-BAMS beads that have different specificities. With Affi-BAMS, both intact proteins and protein fragments can be monitored for targeted proteomics applications.
Affi-BAMS reagents have been created for protein targets in the areas of oncology, neurology, epigenetics, virology, and food allergens. The presentation will introduce the Affi-BAMS workflow and highlight several application areas.
Role of UDP-Glucuronosyltransferases in Drug Metabolism and Drug-Drug Interactions
(Date : 5th Oct 2021)
The importance of glucuronidation for
metabolism of xenobiotics in humans is illustrated by the abundance and broad tissue distribution of the enzymes, the diversity of substrates and reactions catalyzed, as well as a number of distinctive UGT enzymes and their genetic polymorphisms. The involvement of UGTs in metabolism of new drug candidates will be presented as it underscores the enzymes’ importance from regulatory and safety perspectives.
Basic biochemical characteristics of UGTs will be reviewed and examples of major forms of glucuronides formed in humans will be presented. Distinctive properties of acyl-glucuronides and their significance for drug-drug interactions will be highlighted.
Factors contributing to underprediction of UGTs’ in vivo contributions to the metabolism of xenobiotics will also be covered. Laboratory approaches aimed at
improving these predictions will be covered, as well.
Key concepts discussed in this webinar will include:
Significance of UGT-mediated metabolism in drug development process
Consequences of formation of acyl-glucuronides
Underprediction of in vivo UGT-mediated metabolism
ACHIEVE YOUR RESEARCH AND DRUG DISCOVERY NEEDS WITH HTRF ASSAYS
( Date : SEPT 03, 2021 03:00 PM IST )
HTRF no-wash technologies provide a larger dynamic range, improved sensitivity, and higher signal-to-noise ratios than traditional wash-based ELISAs. HTRF has been proven in a variety of applications including GPCRs, kinases, epigenetics, PPIs and the quantification of a wide range of biomarkers including cytokines. Benefit from PerkinElmer’s broad offering of reagent technologies and ditch your ELISA wash steps for good!
ADDRESSING IN VITRO EYE IRRITATION WITH THE SKINETHIC HCE : RECENT DEVELOPMENTS AND LIVE LAB SESSION
(Date : JUNE 01, 2021, 9 AM CET & 4 PM CET )
Validated and integrated in the OECD
TG492 guideline, the SkinEthic HCE model allows to replace the animal to identify products not requiring classification for eye irritation.The webinar will present the characterization of this model, its specificities and the importance of quality control in its manufacturing process to deliver a model with high performance every week.
Its use in the framework of the OECD guidance Document on Integrated Approaches to Testing and Assessment (IATA) for Serious Eye Damage and Eye Irritation for regulatory toxicology will be presented as well as the new perspectives opened by the ongoing validation of the Time to toxicity method to identify mild irritants (category 3).