Cyto-Mine®

The Cyto-Cellect® Mouse Kit from Sphere Bio is designed for use with the Cyto-Mine® Single Cell Analysis Platform to detect and isolate mouse IgG-secreting cells. Using a sensitive FRET-based assay, the kit enables rapid and reliable identification of high-productivity cells for applications such as antibody discovery.

The steps to create a stable, manufacturing cell line include host cell line selection, engineering of the expression vector, transfection, cloning and cell expansion, as well as various screening steps to select the clone with the highest cell viability, growth, expression level, stability and product quality. In this eBook, Dr. Jon Dempsey discusses his thoughts regarding best practices for optimizing cell line development (CLD) processes.

Dr. Jon Dempsey is the CEO and Founding Partner of Pathway Biopharma. Dempsey has spent nearly 30 years working in the Biopharmaceutical industry, bringing expertise in biologics chemistry, manufacture and control (CMC) including cell line development, cell therapy and process development, analytics and protein chemistry, CDMO selection and manufacturing. During this time, Dempsey has been involved in the development of several commercial biotherapeutics. In this eBook, Dempsey shares advice for those researchers/ scientists starting out and for those currently in CLD, including tips on the best strategies for process improvement.

In this application note, Sphere Fluidics Ltd., presents a method by which Cyto-Mine®, utilizing an antigen-specific assay, analyses a heterogeneous B cell or hybridoma population to identify target-specific variants in a high-throughput manner. The Cyto-Mine Single Cell Analysis and Monoclonality Assurance System is the first fully integrated platform designed specifically for biopharma that screens millions of primary B cells or hybridomas for secreted immunoglobulin, identifies and sorts the antigen-specific candidates, and then gently dispenses them into 96- or 384-well microtiter plates with visual proof of cell number, through high-quality imaging.

In the discovery and development of novel biopharmaceuticals (antibodies) for therapeutic use, cell line development is an integral process in the biomanufacturing of therapeutic proteins. With the market demand for biotherapeutics increasing at a dramatic rate there is a pressing need to substantially shorten the development workflow. Cyto-Mine® utilizes established picodroplet technology to fast-track the generation of cell banks comprised of high-producing clones derived from single cells with monoclonality assurance. In this application note, Sphere Fluidics illustrates the process by which the Cyto-Mine® IgG secretion assay measures the productivity of hundreds of thousands of single cells encapsulated in highly consistent picoliter droplets or ‘test tubes’ while simultaneously providing verification of clonality.

Over the last few years, there has been an increasing interest in cell therapy as a strategy to harness the immune system to fight tumors. Technologies include reprogramming of T cells collected from patients to produce personalized medicines. T-cell therapy capitalizes on the body’s immune systems ability to recognize and kill tumor cells. This ability can be lost or overwhelmed as a cancer develops. By extracting and manipulating a patient’s T-cells, the ability to attack cancer cells can potentially be restored. Rapid, accurate assays to qualify the potency of the engineered T-cells against the target tumor cell are required to support the effective and efficient development of therapeutics. In this application note, Sphere Bio presents a new picodroplet-based approach for the functional validation of CAR-T cells. The easy-to-use and robust method combines the benefits of Sphere Fluidics’ high-throughput picodroplet technology with a granzyme B assay for studying cell-mediated cytotoxicity. Engineered T cells and target cells are co-encapsulated in picolitre-volume aqueous droplets (picodroplets) in an oil emulsion, along with granzyme B substrate. Granzyme B release by the T cell, a readout of the cells ability to recognize and kill the cancer cell, can be captured to determine the level of potential anti-cancer activity. In doing so, this method enables rapid cell product efficacy readouts to streamline the QC release of cell product.

Picodroplet technology is a transformative single cell analysis technique based on the rapid encapsulation of individual cells in miniaturized, picoliter-volume, water-in-oil droplets (picodroplets). Each picodroplet serves as a miniature test tube, providing the ability to screen single cells in their own microenvironment with increased precision and sensitivity. This format provides a uniquely protective environment to support cell viability and integrity during incubation, and shield cells against shear stress during subsequent analysis. In this application note, Sphere Bio demonstrates the feasibility and flexibility of using picodroplet technology for applications that involve prolonged incubation times. This technique can be used for a wide range of applications including identifying single cells that produce the highest antibody concentrations, isolating rare cells from large populations, and undertaking genome-editing analysis at the single cell level.

Picodroplet microfluidics is rapidly becoming the technology of choice for single-cell analysis, providing an easy-to-use tool to encapsulate single cells in picolitre-volume aqueous droplets (picodroplets) at high throughput rates of thousands of droplets per second. This powerful and sensitive technique offers significant cost and efficiency advantages. An additional advantage of this approach is that picodroplets provide a cell friendly culturing environment. Depending on the assay and application of interest, cells can be cultured in these picodroplets for a couple hours, to up to several days. The ability to culture the cells for various lengths of time has opened opportunities for a wide range of applications including antibody discovery, cell line cloning and cell engineering. In this application note, Sphere Bio provides an overview of how picodroplets can be used in cell engineering.

Dr Fay Saunders, UK Head of Upstream Mammalian Cell Culture, Process Development at FUJIFILM Diosynth Biotechnologies, shares her experience of using Cyto-Mine® in cell line development and recombinant protein expression.

This application note will demonstrate how Cyto-Mine® can accurately identify and isolate ‘hit’ cells using an antigen-specific assay from a large hybridoma population, quantify varying concentrations of antigen-specific antibodies, and significantly shorten the antibody discovery workflow by using a single fully integrated instrument.

Cutting-edge technologies and collaborative research are transforming how scientists overcome genomic instability and accelerate therapeutic protein production

The name change supports the company’s global commercial growth, driven by customer demand for integrated life sciences tools and solutions

Discover how droplet microfluidics is revolutionizing single-cell analysis and selection in biopharma

From monoclonality to droplet-based technology, find out the key to cell line development success

In this on-demand webinar, Dr. Devika Kalsi, and Dr. Xin Liu illustrate how they can reduce cloning timelines from 25 to 10 weeks

Simplified design reduces manufacturing time and offers improved functionality to help streamline antibody discovery, cell line development, cell therapy, and synthetic biology workflows

Partnership awarded Innovate UK Knowledge Transfer Partnership grant

Single-cell analysis for therapeutic discovery recognized with prestigious business award for innovation

Pico-Surf is designed to offer reliable, highly stable and high-quality droplet generation and processing for microfluidic applications

Integration of S-LAB with the Cyto-Mine Single Cell Analysis System increases throughput in antibody discovery and cell line development