Automated single plasma B-cell secretion screening and recovery
Accelerating antibody discovery: Parallel analysis and isolation of thousands of single plasma B cells within one day
The 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.
Our new method for the analysis and isolation of single B-cells uses the CellCelector™ together with the unique Nanowell technology and combines selective cell screening, imaging, sensitive single cell assays as well as cell isolation into a single platform. Single cell analysis allows also the detection of rare antibodies with unique properties which are hard to find under conventionally 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 the plate type as well as the size of the nanowells, this results in between 60,000 and 200,000 nanowells per well. After single cell solution is added, the individual cells are randomly distributed in the nanowells according to the Poisson distribution.
H 100 array: hexagonal 100 µm nanowells seeded with B-cells and antigen-specific coated beads
Step 2: Single cell screening
The seeded wells are scanned with the ALS CellCelector™ in bright field. The software automatically identifies single cell nanowells.
Step 3: Running the secretion assay
The specific secretion assays are performed in the nanowells on the individual cells and allow the rapid identification of secreting cells of interest.
The CellCelector™ B-cell workflow supports several types of cellular assays, e.g.
Bead-based antigen specificity assay
The plasma B-cell are seeded together with antigen-specific coated beads.
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.
Antigen-expressing reporter cell assay
The plasma B-cells are seeded together with reporter cells expressing specific antigen.
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), the ALS CellCelector™ transfers them to the appropriate target vessels for subsequent analysis and processes.
The ALS 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 a later review. 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 LIMS system.
Advantages of the CellCelector™ single plasma B cell workflow
Robustness
- No fusion step required
- No B-cell loss
- No cross-contamination between recoveries due to single-use capillaries
Sensitivity
- Extremely gentle cell transfer guarantees healthy and vital B-cells after screening and sorting for use in downstream applications
Flexibility
- CellCelector
- In depth characterization: affinity, specificity and functionality
Automation
- Full automation of the complete process from cell loading to export of hits with full documentation of the whole screening and cell recovery process
Precision
- Exact identification of single cells in subnanoliter volume of nanowells
- High image quality up to 40x magnification
Throughput
- Screening of thousands of single B-cells in parallel thanks to CellCelector
Efficiency
- Screening process shortened from several months to one day
- Increased success rate for discovering rare antibodies
- Considerable time and cost savings; affordable consumables
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