サポート ブログ Understand Single B Cell Antibody Technology in One Read

Understand Single B Cell Antibody Technology in One Read

Biointron 2024-03-15 Read time: 5 mins

Antibody drugs are a growing force in the biopharmaceutical market, thanks to their precise targeting, powerful therapeutic effects, and favorable pharmacokinetics. As demand for these drugs increases, so too does the need for efficient methods to discover and develop new antibodies. Several technologies exist, including hybridomas, phage display, and single B cell cloning. While each has its strengths, this article focuses on the promises of single B cell antibody technology.   

1. What is single B cell antibody technology?

Single B cell antibody technology harnesses the natural power of B cells. B cells can produce specific antibodies from immunized animals, which contains a functional heavy chain variable region DNA sequence and a light chain variable region DNA sequence that can be directly amplified. The scientists isolate individual B cells and retrieve the genetic blueprints for specific antibodies.  

This technology has revolutionized antibody discovery and is now widely used in research targeting infectious diseases, tumors, autoimmune conditions, and organ transplantation.  

2. What are the advantages of single B cell antibody technology?

Traditional hybridoma technology has limitations, including a lengthy development cycle that can take months due to immunization steps. Additionally, the fusion process may only capture a small portion of B cells, hindering comprehensive screening of the vast antibody library. Phage display boasts a large library capacity, but also has drawbacks. The pairing of heavy and light chain variable regions (VH and VL) is random, and can result in combinations that may not function optimally. 

Single B cell antibody technology emerges as a next-generation solution offering several key advantages: 

  • Faster timeline 

  • More positive clones 

  • Can be a fully human source 

  • Small amount of cells required 

Most importantly, single B cell technology preserves the natural pairing of light and heavy chains, which is necessary for optimal antibody function, leading to potentially more potent and specific therapeutic candidates.  

3. What is the process of single B cell antibody technology?

Single-B-Cell-Workflow.jpg

Here's a breakdown of the key steps involved: 

  1. B Cell Enrichment: After exposure to a target antigen (like a virus or protein), B cells that produce the desired antibody are derived from peripheral blood collection. 

  2. Single B Cell Screening: The most important step of the process, individual B cells are isolated and evaluated to identify those actively producing the antibody of interest. Microfluidic-based methods can achieve higher throughput and greater flexibility over nanowell and nanovial techniques. 

  3. Antibody Gene Sequencing and Analysis: Once a promising B cell is identified, scientists can use single-cell PCR to amplify and sequence the genes encoding the antibody's heavy and light chains. 

  4. Recombinant Antibody Expression: The isolated antibody genes are introduced into host cells, which are then engineered to produce large quantities of the specific antibody. Mammalian host cells are the most commonly used. 

  5. Antibody Function Verification: The functionality and specificity of the produced antibody are rigorously tested to ensure it meets the desired criteria, such as with flow cytometry, enzyme-linked immunosorbent assay, and immunohistochemistry. 

4. Development of a high-throughput antibody discovery platform based on single B cell technology

Biointron's high-throughput single B cell screening platform can screen 2x106 plasma B-cells from immunized animals. Antigen-specific antibody-secreting cells can be isolated, allowing for the detection of potentially rare antibodies and drugs. Millions of cells can be screened within 1 week using single-cell droplet generation and sorting. Learn more by reaching out to us at info@biointron.com.

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