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Fluorescent Activated Cell Sorting (FACS) Technology for Single Cell Isolation

After years of experience in antibody discovery and manufacture, Creative Biolabs has established a distinguished Native™ Antibody Discovery platform. We are proud to offer mAb discovery service based on fluorescent activated cell sorting (FACS) technology in a timely and cost-effective manner.

Native™ Antibody Discovery platform, also known as single B cell antibody discovery technology, is thought to be an important and efficient strategy to generate antibodies, which allows the direct amplification of VH and VL region encoding genes from single B cells and their subsequent expression in cell culture systems. Creative Biolabs offers a wide range of strategies for B cell isolation. FACS has become one of the most accepted, worldwide popular strategies in identification and selectin of B cells.

FACS helps to perform random B cell isolation and antigen-specific B cell selection based on the expression patterns of specific cell surface markers. For antigen-specific B cell selection in native antibody development, multi-parameter FACS is commonly used by combining different fluorochrome-labeled markers and antigens or antigen coated magnetic beads and fluorochrome-labeled antigens. By using FACS, B cells in different stages of development and differentiation can be clearly distinguished.

Fluorescent Activated Cell Sorting (FACS) Technology for Single Cell Isolation Fig 1. Schematic overview of single-cell flow cytometry separation technologies. (Gross, 2015)

Workflow of native antibody discovery by FACS

To meet our customers' requirements, Creative Biolabs can provide antibody discovery service based on FACS. Briefly, single B cells are isolated from PBMCs, lymph nodes or spleen of individuals by Ficoll separation or magnetic B-cell enrichment. Single B cells to specific antigen are identified by staining with the biotinylated or fluorescently labeled antibodies followed by sorted into 96-well or 384-well PCR plates. Single-cell cDNA synthesis and gene amplification are performed. Meanwhile, detailed Ig gene sequence of all PCR products are analyzed. To verify the characteristics of antibodies or fragments, they are produced by bacterial expression systems (e.g. Escherichia coli) or transient and/or stable mammalian cell systems (e.g. HEK 293, CHO cells) with IgH and IgL expression-vectors. The antibodies are affinity purified and then finally characterized in details at a molecular and functional level.

Fluorescent Activated Cell Sorting (FACS) Technology for Single Cell Isolation Fig 2. One strategy of Ab generation by FACS.

Features of single cell isolation via FACS


Based on every client’s requirement, Creative Biolabs provides custom strategies for your native antibody development. Please don’t hesitate to contact us if you require more information.

Reference

  1. Gross, A.; et al. Technologies for single-cell isolation. Int J Mol Sci. 2015, 16: 16897-16919.



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