Accelerating Antibody Discovery: Parallel Analysis and Isolation of Thousands of Single Plasma B Cells within 8 hours
Identification and selection of optimal clones that secrete high levels of the desired antibody is one of the critical and time-consuming steps in the antibody discovery process.
CellCelector - together with the unique Nanowell technology combines selective cell screening, imaging, sensitive single-cell assays and cell isolation into a single platform. Single-cell analysis also allows detection of rare antibodies with unique properties which are hard to find under conventional screening methods. Individual cells will be immediately tested in several assays (instead of culturing for weeks) to reach a minimum number of cells for the assay.
CellCelector One Day B Cell Screening and Isolation Workflow
Step 1. Easy Cell Seeding
The single-cell suspension is dispensed manually into the wells of the nanowell array. Each well of the nanowell array is characterized by an additional well pattern on its bottom. Depending on plate type and the size of the nanowells, this results in between 60,000 and 200,000 nanowells per well. After the single cell solution is added, the individual cells are randomly distributed in the nanowells according to the Poisson distribution.
Figure 1: H 100 array: hexagonal 100 µm nanowells seeded with B cells and antigen-specific coated beads
Step 2. Single Cell Screening
Seeded wells are scanned with the CellCelector Flex in bright-field. The software automatically identifies single-cell nanowells
Step 3. Running the Single Cell Secretion Assay
Antigen Specificity Assay with Plate Capture Coating
The surface of the nanowell array is coated with antigen and blocked with BSA solution before plasma B-cell seeding.
The Specific secretion assays are performed in the nanowells on the individual cells to allow for the rapid identification of secreting cells of interest. The CellCelector B-cell workflow supports several types of cellular assays.
Figure 3a: Assay with plate capture coating. The surface of the nanowell array is coated with antigen and blocked with BSA solution before plasma B-cell seeding.
Antigen-Expressing Reporter Cell Assay
The plasma B-cells are seeded together with reporter cells expressing specific antigen.
Figure 3b: Antigen-expressing reporter cell assay
Bead-based antigen specificity assay
The plasma B-cell are seeded together with antigen-specific coated beads.
Figure 3c: Bead-based assay
Other Possible Assays:
- Receptor/ligand blocking assay
- Cell surface marker assay
- Multiplexed secretion assay
- Functional assay (e.g. GFP-expressing cells)
Step 4. Cell Transfer
After identification of the desired cells (hits), CellCelector transfers them to the appropriate target vessels for subsequent analysis and processes. The CellCelector facilitates a very gentle picking process based on mechanical aspiration, resulting in high rates of viability of the picked B cells.
Step 5. Generation of the Report
The entire process from single-cell screening to transfer of the hits is fully documented and compliant to GLP and GMP standards. High-quality microscopic images of each picking process allow for review at a later time. All imaging and numeric data (e.g., morphology and spectral values) as well as source and destination positions of each cell are stored in the database and can be exported quickly and easily into the customer’s LIM system.
