Biomarker Detection Reagents

The main vectors of gene therapy in research are viruses. The most popular tool for gene delivery is a genetically modified lentivirus. Modified lentivirus (HIV-1) vectors retain their ability to infect undivided cells, thereby increasing their ability to transduce a wide variety of cells, including those that are difficult to transduce. This advantage enables the stable long-term expression of a transgene. In immunotherapy, CAR-T cells are manufactured by transducing the CAR gene with an HIV-1 vector in T cells to express a specific chimeric p24 protein on their surface. This allows them to recognize cancer cells and destroy them. These CAR-T cells must be generated individually to treat each patient. This application note demonstrates a comparative quantification of the p24 titer in a lentiviral GFP control sample using Alliance HIV-1 p24 Antigen ELISA and p24 AlphaLISA immunoassay platforms. Check out the different sections of this application note: A lentiviral vector that encodes the CAR construct Determination of the efficiency of transient co-transfection by measuring viral titer Detection of the presence of a p24 HIV-1 specific antibody in the sample Quantification of targets present in the sample

Non-alcoholic fatty liver disease (NAFLD) describes a progressive pathology that affects the liver. Fat accumulation causes fatty liver (NAFL) or steatosis to develop, which leads to lipotoxicity and in turn induces liver inflammation and apoptosis, resulting in non-alcoholic steatohepatitis (NASH). NASH can progress to fibrosis and then cirrhosis, which in some cases will lead to hepatocellular carcinoma (HCC). Read this case study to learn how non-invasive preclinical in vivo imaging was used to longitudinally visualize, quantify, and diagnose NASH with the goal of investigating the efficacy of liver fibrosis-preventing drugs on NAFLD progression.

What Are the Challenges and Solutions? All across the globe, AAVs are getting the attention of scientists and companies working in gene therapy, as they provide the right combination of characteristics that make them one of the most promising vector today. In 2021 only, the global gene therapy market was 4.1 billion USD, and AAV vector therapies made up more than 43% of that market value! Designing and manufacturing AAV vectors is complex, and to be successful, certain challenges must be addressed. This implies monitoring and optimtizing production for a thorough quality control process, including reliable ongoing characterization of process intermediates and the final product. Key features include: Overview and data on the gene therapy market Description of AAV vectors’ genomes and transfer Challenges associated with AAV vectors’ design and manufacturing Analytical methods for AAV vector quality control with a description of innovative no-wash AlphaLISA™ assays

In the dynamic landscape of pharmaceuticals, biotherapeutics research stands out as the fastest-growing sector. Scientists worldwide are on a quest for ever more efficient tools to develop therapeutic proteins. From lead selection to bioprocessing qualification, rigorous analytical characterization is essential. That’s where Revvity steps in. Our comprehensive line of no-wash tools revolutionizes biologics screening, mechanism-of-action studies, and biomanufacturing. With robustness and convenience at the forefront, we empower researchers to accelerate discoveries and drive innovation. Welcome to a new era of precision and efficiency in biotherapeutics research. For research use only. Not for use in diagnostic procedures.

Detecting and quantifying HCPs with automatable homogeneous immunoassays During biotherapeutic manufacturing and production, the host cells - a great majority of them being Chinese Hamster Ovary (CHO) cells – produce protein impurities that are called Host Cell Proteins (HCPs). Even if more than 99% of them are removed from the final product, the residual CHO HCPs can induce immunogenicity in individuals or reduce the potency, stability, or effectiveness of a drug. Therefore, to meet regulatory organizations’ guidelines (such as FDA or EMA) on CHO HCP levels, biopharmaceutical companies spend significant amounts of money on tools and strategies for their detection. Illustrated with robust results, this Application Note explains the many ways in which HTRF™ and AlphaLISA™ kits can improve the workflow for CHO HCP detection: Wide antibody coverage Compatibility with most commonly used buffers No cross-reactivity between CHO HCP detection and drug substance Good dilutional linearity and antigen spike recovery

Alpha technolgy enables the rapid and straightforward mesaure of virtually any target. This includes enzymes, receptor-ligand interactions, low-affinity interactions, second messenger levels, DNA, RNA, proteins, protein-protein interactions, peptides, sugars and small molecules. Explore the vast selection of homogenous Alpha assays and reagents, including AlphaLISA ™ , AlphaLISA ™ SureFire ® Ultra ™ , AlphaScreen ™ , and Alpha Toolbox. For research use only. Not for use in diagnostic procedures.

Discover Alpha SureFire ®   Ultra ™ assays, the no-wash cellular kinase assays leveraging Revvity's exclusive bead-based technology and sandwich immunoassays for detecting phosphorylated proteins in cells. Offering a quantitative alternative to Western Blotting, Alpha SureFire assays are automation-friendly, easily miniaturized, and proficient in detecting both endogenous and recombinant proteins. Explore our comprehensive portfolio of SureFire Assays, designed to help you elevate and expedite your drug discovery journey.

Orthogonal systems to cell-based assays are a key requirement in EMA/FDA guidelines for potency estimations and require cross-validation with complementary approaches to prove and strengthen the reliability of results. In this application note published in collaboration with IBR Inc., you will learn: Why Alpha technology represents an ideal cell-free orthogonal system for potency assays How AlphaLISA assays can be used to determine Bevacizumab/VEGF165 potency An example of how to run an AlphaLISA potency assay and the type of data that can be generated Why it is suitable for assessing lot-to-lot consistency and equivalence of Bevacizumab and biosimilars

This guide outlines further possible optimization of cellular and immunoassay parameters to ensure the best possible results are obtained.

The definitive guide for setting up a successful AlphaLISA SureFire Ultra assay Several biological processes are regulated by protein phosphorylation. It is, therefore, no surprise that the dysregulation of protein phosphorylation is implicated in a relatively large number of diseases. AlphaLISA SureFire Ultra assays provide a robust and reliable method for quantifying a targeted phosphorylation event in cell-based experiments. This guide contains tools and data helpful for you to perform your assays using AlphaLISA SureFire Ultra: A detailed description of the assay and its options A thorough investigation of assay conditions to obtain the optimal response from the chosen modulator and cell line A list of optimization steps to provide a sufficient assay window and produce the strongest results possible

The AlphaPlex™ reagent system was designed to enable fast and easy transition from well-established AlphaLISA™ assays to multiplexed detection of a broad range of proteins, molecules and biomarkers. Using a universal, streptavidin-coated Donor bead, multiple AlphaPlex Acceptor beads targeted to various analytes are combined in a single assay well. Download this guide to learn how to easily set up your AlphaPlex assay and streamline your workflow! For research use only. Not for use in diagnostic procedures.

Atherosclerosis pathogenesis, cellular actors, and pathways Atherosclerosis is a common condition in which arteries harden and become narrow due to a build-up of fatty material, usually cholesterol, and other substances such as calcium. This can lead to a range of serious health complications, including heart attack or stroke, making the disease an important contributing factor in death and morbidity in developed countries. Recent developments in our understanding of atherosclerosis from a molecular perspective include the discovery of new players in disease pathogenesis. Included in this white paper Atherosclerosis: step-by-step pathogenesis, therapeutic strategies, and recent developments Detailed descriptions and explanations, including a focus on pathways

With over 200 different types of cancer, management relies on a variety of techniques such as chemotherapy, radiotherapy, and surgery. These types of therapies can be associated with severe side effects, and finding safer and more effective treatments is a priority in cancer therapeutics research. One approach that has shown huge potential is monoclonal antibody-based cancer therapeutics. In this white paper we explore this exciting area of anti-cancer research, covering mechanism of action, development, and challenges in monoclonal antibody-based therapeutics, including antibody-drug conjugates and multispecific antibodies.

Leptospirosis is an infectious disease caused by Leptospira bacteria, which can spread across different species of mammals. Transmission occurs through contact or consumption of food from infected domestic animals. In cattle, Leptospira infection leads to reproductive failure, affecting productivity and profitability in the dairy and beef industries. Vaccinations are widely used to prevent infection in animals and reduce transmission to humans. This application note: demonstrates the performance of AlphaLISA bovine cytokine kits by measuring the cytokine release from antigen stimulated PBMCs isolated from cow blood shows how cytokine measurements are critical for understanding cell mediated immune responses during vaccine development.

Immunoassays are a mainstay for the quantification of a variety of biomolecular analytes in drug discovery, drug development, and life sciences research. AlphaLISA® proves advantageous to ELISAs, offering a novel, homogenous immunoassay technology that eliminates wash steps. Using the JANUS® Automated Workstation in combination with AlphaLISA provides a solution to easily preparing assays that can be tailored to the needs of the laboratory. Moreover, the JANUS workstation can be easily set-up to process different assay types in a multi-user, multi-assay environment, thus providing flexible automated workflows.

AlphaLISA™ technology is a highly sensitive, easy-to-use, and reproducible method for detecting and quantifying molecules in various biological matrices. It works by using streptavidin-coated Donor beads and biotinylated anti-analyte antibodies. When these come into close proximity, the excitation of the Donor beads at 680 nM triggers an energy transfer cascade in the Acceptor beads, generating a sharp emission peak at 615 nM. However, some cell culture media contain high levels of biotin, which can interfere with AlphaLISA and other assay technologies that rely on a streptavidin-biotin binding event for detection. High levels of free biotin in the sample matrix can result in a decrease in total counts, lower signal to background ratios, and reduced AlphaLISA assay detection limits. To mitigate this, AlphaLISA biotin-free kits have been developed. This application note demonstrates the value of using AlphaLISA biotin-free kits to reduce the effects of biotin interference in sample and standard preparations.

Fluorescence molecular imaging is the visualization of cellular and biological function in vivo to gain deeper insights into disease processes and treatment effects. Designing an effective study from the beginning can help save time and resources. Learn about several important best practices, from proper probe selection to study design to imaging technique tips and tricks needed to generate meaningful biological information from your in vivo fluorescence imaging studies.

Cytokine Detection Using AlphaLISA Biotin-Free assays This technical note describes the use of AlphaLISA biotin-free kits for detecting and quantifying cytokines, including interleukin 2 (IL-2), tumor necrosis factor alpha (TNFα), interferon gamma (IFN-γ), and interleukin 6 (IL-6), in supernatants produced by human PBMCs cultured for two days with or without Dynabead ® stimulation.

The measurement of protein phosphorylation is a useful tool for measuring the modulation of receptor activation by both antibodies and small molecules. CCR7 and CXCR2 receptors, which are expressed in immune cells and are therapeutic targets for disorders like lupus erythematosus, adult leukemia, lymphomas, chronic obstructive pulmonary disease (COPD), and sepsis. AlphaLISA ™ SureFire ® Ultra ™ and Alpha SureFire ® Ultra ™ Multiplex assays are automation-friendly, applicable to both small and large-scale screens, and can assess phosphorylation status in complex matrices. The LANCE Ultra cAMP assay is measures cyclic AMP (cAMP) produced upon modulation of adenylyl cyclase activity by G-protein coupled receptors (GPCRs). This application note demonstrates how the SureFire Ultra and LANCE Ultra cAMP assays can be used for measuring inhibitors to CCR7 and CXCR2 cell surface receptors using a cellular model system where these receptors are overexpressed in CHO cells. The assays were optimized to measure receptor blockage and assayed receptor activity modulation by detecting ERK and AKT phosphorylation status and cAMP modulation. For more details, download the application note!

AlphaLISA and LANCE (TR-FRET) biomarker assays can be used to measure ECM-associated protein modulation caused by human transforming growth factor-beta (TGF-β) induction of EMT in a 3D Spheroid model of human prostate carcinoma.

With the increased understanding of molecular and cellular medicine, more specific and efficient gene transfer vectors are now producing clinical results. 20 years after the first clinical trials, gene therapy is considered one of the greatest scientific success stories of the 21st century. While the Covid-19 crisis has disrupted the evolution in this promising field, gene therapy is expected to recover and continue its growth in the future. Download this complete industry report to learn more about the gene therapy market trends and latest advances. It includes: Gene therapies in the global pipeline and major actors in the field A description of gene therapy development and vectors used The challenges concerning gene therapy manufacturing The promising future of gene therapy

DELFIA® immunoassays are particularly well suited for discovery of specific, high-affinity monoclonal antibodies (mAbs), especially at the low concentrations that may be present in hybridoma supernatants. An assay used in these conditions must be sensitive, specific, reliable, reproducible and easy enough to handle 1,000 to 1,500 samples in one batch.

DELFIA immunoassays are a superior alternative to traditional ELISAs

Cytokines are implicated in the pathology and outcome of disease states across all therapeutic areas. When preparing to run cytokine assays, it is key to consider a multitude of factors that impact the assay performance, such as sample dilutions and cell culture format. Utilize this guide to optimize your AlphaLISA ™ cytokine assays with in-depth data and information!