Single Plasma B-Cell Screening and Picking | Sartorius

Automated Single Plasma B-Cell Secretion Screening and Recovery

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.

Advantages of CellCelector for Single Plasma B Cell Workflow 

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Automated

Screen up to 5 million B-cells or hybridomas in a single plate using Nanowell technology – in less than 8 hours

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Precise

Antigen specific identification and isolation of single B cells

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Versatile

Use of plate coating, beads or reporter cells to capture the antibody of interest

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Flexible

Multiplex screening of up to 14 antigens in a single run

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Efficient

High-throughput interaction studies of multiple cell types in parallel

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Resources

Featured Resources

eBook

The Future of Antibody Discovery for Biotherapeutics

Rapid, automated workflows for screening, isolating, and assaying antibody-secreting cells. enabling researchers to assay thousands of single primary...

Application Guide

CellCelector Flex for Antibody Discovery and Production

Fully automated, image-based screening and isolation of clones and single cells for antibody discovery.

Biologics Molecule Development Workflow - Sartorius Solutions for Every Step of the Process infographic

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