Antibody Discovery

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.

Fluorescence-Activated Cell Sorters (FACS), while faster and higher throughput than limiting dilutions, operate at high system pressure (up to 70 psi) that can easily damage or kill the sorted cells. Induced pluripotent stem (iPS) cells, a commonly used cell type for CRISPR cloning, are particularly sensitive to the high pressure and have low survival rates following isolation. Additionally, FACS is difficult to operate and prone to contamination.

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.

Namocell Benefits

Gentle

Gentle sorting (< 2 psi) preserves cell viability Higher clonality, better outgrowth

Easy

Fit inside a cell culture hood for sterile sorting No clogging, no cross-contamination; Easy to use, zero maintenance

Fast

Quickly isolate rare antigen-specific B cells Process 1M cells in 5 min

Dispensing Efficiency

Namocell Single Cell Dispensers are highly efficient and consistent at dispensing single cells (80-90% with mammalian cells) compared to manual pipetting (~30%).

Single B Cell Isolation

Namocell’s Namo 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.

What Scientists are Saying