Monoclonal antibodies are essential for modern drug development and diagnostics. Scientists have traditionally identified antigen-specific monoclonal antibodies with two approaches, each with significant drawbacks that limit the efficiency of antibody discovery.
Screening for hybridoma clones. This entails isolating B cells and fusing them with immortalized myeloma cells. Traditionally, limiting dilutions are used to isolate the desired clones. However, limiting dilution is inherently limited by the Poisson distribution, making this approach highly labor-intensive and time-consuming.
Screening for antigen-specific immune cells. Antigen-specific B cells can be directly isolated by FACS instruments, which sort cells at very high pressure (up to 70 psi), resulting in reduced cell viability. Additionally, these instruments are very difficult to operate and prone to clogging.
Two methods for monoclonal antibody production are depicted above: the creation of immortalized myeloma cell lines (left), and a screening process to isolate antigen-specific B-cells that produce the desired IgG antibodies (right). In both workflows, Namocell’s Single Cell Dispensers are used to isolate single cells for downstream analysis.
- Gentle sorting (<2 psi) preserves cell viability
- Higher clonality
- Better outgrowth
- Quickly isolate rare antigen-specific B cells
- Process up to 1M cells in 5 min
- Quick setup in under 3 min
- No calibration and no maintenance
- Compact size fitting in tissue culture hood
- Sterile and disposable cell cartridge
Namocell Single Cell Dispensers are highly efficient and consistent at dispensing single cells. With dispensing efficiency (percentage of dispensed wells with single cells) of 80-90% for CHO cells compared to ~30% using manual pipetting.
Namocell’s Single Cell Dispenser was used to isolate single memory B cells for cDNA library construction. Expression of immunoglobulin genes via qPCR confirmed the uniform presence of heavy chain IGVH and the presence of either the kappa or lambda light chains (IGVK, IGVL) in eight representative single cells.
High-throughput and automation advances for accelerating single-cell cloning, monoclonality and early phase clone screening steps in mammalian cell line development for biologics production. Tejwani V, et al. Biotechnol Prog. 2021 Nov;37(6):e3208. doi: 10.1002/btpr.3208. Epub 2021 Sep 20. PMID: 34478248.