Primary B-Cell Analysis
Antibody-derived biologics have become a major class of therapeutic, particularly in the fight against cancer and autoimmune diseases. Recent developments in single cell gene sequencing and recombinant immunoglobulin expression have enabled direct screening of native B-cells (B-lymphocytes), giving the potential for much more of the mammalian immune system repertoire to be deep-mined.
Current techniques such as cell sorting are poor at analysing the antibody secreted from each cell. The process can be far more effectively streamlined with our high-throughput picodroplet technology. Screen greater numbers of cells in a shorter amount of time and don’t miss finding rare and valuable variants within your B-cell repertoire.
The Unique Benefits of Our Systems for Primary B-Cell Screening
Using our microfluidic technology for B-cell analysis involves animal immunisation and the subsequent isolation of B-cells. What’s the key difference? By using picodroplets, we are able to analyse whole B-cell repertoires (tens of millions of B-cells) in a single run and isolate and dispense all positively secreting B-cells whether just by total IgG production or antigen-specific assays. Once these cells have been identified, isolated and dispensed, they can be subject to further downstream processing e.g. further assays, RT-PCR of antibody transcripts, next generation sequencing etc.
Our Systems in Action for Primary B-Cell Screening
We understand the importance of maintaining cell viability throughout the research process, particularly when working with sensitive cells. Our technology ensures that the viability of primary B-cells remains high when encapsulated and analysed in picodroplets. As can be seen in the graph, cells incubated in 300 pL picodroplets remain viable over the four hour incubation period that is standard for most of the workflows carried out using our systems. This viability is crucial in primary B-cell analysis, as the cells often need to be happy and healthy after screening and sorting, so they are ready for use in downstream applications.
Primary B Cell Analysis:
Recent developments antibody drug discovery, through single cell gene sequencing and recombinant immunoglobulin expression, have enabled direct screening of native B cells (B lymphocytes), giving the potential for much more of the mammalian immune system repertoire to be deep-mined. Cyto-Mine® is a unique platform which allows you to rapidly screen vast numbers of native B cells to find rare cells of interest.
Hybridoma generation and screening is a powerful tool for the discovery of novel monoclonal antibodies. Current techniques used in the field are restricted by the number of hybridomas they are able to screen each day. This can reduce the quality of hits obtained and slow down development pipelines.
Our high-throughput picodroplet technology can rapidly screen your hybridoma library, helping you to isolate highly-viable cells producing high quality antibodies against your target antigen, ready for further downstream analysis.
The Unique Benefits of Our Systems for Hybridoma Screening
Efficient hybridoma screening is essential to ensure you’re generating high-quality, antigen-specific antibodies for use in research, biopharmaceuticals and diagnostics. Using traditional screening methods can be extremely time-consuming and laborious. Clone pickers for example, are able to screen 10,000 hybridomas in 3 to 6 weeks, whereas our picodroplet technology can screen several million hybridomas a day.
The unique speed of our systems allows you to test your entire library and therefore improve the quality of the antibodies you select. Using this miniaturised screening approach enables you to isolate clones that are the top antibody expressers and measure genetic drift in a population. The latter will enable you to more rapidly observe the potential emergence of no or low-antibody expressing sub-populations. Our integrated and compact systems also offer further cost savings, as they reduce the continual spend on screening consumables by more than 50-fold.
We understand the importance of maintaining cell viability when screening for hybridomas. As seen in the graph, during incubation over four hours, viability of the hybridoma cell line is maintained at over 80% to ensure a high number of cells are available for further testing.
Our Systems in Action for Hybridoma Screening
To demonstrate how effective picodroplets are for screening hybridomas and isolating antibodies, we measured monoclonal antibody secretion by single cell hybridomas in picodroplets. The cells were incubated at 37°C over the course of three hours and antibody secretion measured using a FRET-based ELISA (i.e. a homogenous assay). As can be seen in the graph, cells grown in picodroplets performed as well as cells grown in bulk culture over the time period analysed. This allows users to screen a huge number of hybridomas in a short time frame with minimal use of reagents, all without compromising on assay performance.
Application Note: High-Throughput Method for Mining B Cell Repertoires and Hybridoma Fusions to Find Rare Cells Secreting Antigen-Specific Antibodies
Discover how Cyto-Mine® screens complex populations of primary B cells and hybridomas using a miniaturised, high-throughput, homogeneous secretion assay to analyse and identify antigen-specific clones for subsequent sorting and collection to microplates.