Single B Cell antibody discovery technology

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• Monoclonal antibodies (mAbs) are important tools for many diagnostic applications and research.  In addition, monoclonal antibodies have become an ideal therapeutic product for many diseases, and have been well used in the treatment of cancer, autoimmune diseases and infectious diseases.   A number of techniques are used to generate monoclonal antibodies for research and therapeutic purposes.  Single B cell screening is a newly developed technique for rapid preparation of mAbs in recent years. The principle is that each B cell only contains a pair of functional heavy and light chains, and each B cell only produces a specific antibody characteristic, which can be directly amplified from a single B cell to obtain the mAb. This method has the advantages of high speed, high throughput, and natural pairing of variable regions of antibody weight and light chains. It is one of the new and efficient methods for antibody discovery.

• Process of Single B Cell Antibody Production

  

• The Concept of Single B Cell Antibody discovery technology

  The principle of single B cell antibody discovery technology is that each B cell contains only a pair of functional heavy chain and light chain and each B cell has the characteristic of producing only one specific antibody.  Therefore, the antibody coding sequence can be amplified directly from a single B cell to obtain a monoclonal antibody.

• Single B Cell Sorting

  Depending on the research application, single B cells can be isolated randomly or antigen-selectively from peripheral blood or lymphoid tissues (bone marrow, spleen).  For the isolation of B cells, current single B cell sorting technology include flow cytometry, magnetic bead cell sorting, micromanipulation, laser microdissection, and microfluidic sorting.  Fluorescence activated Cell Sorting (FACS) is a mature and effective single cell sorting technology.

  

• Single B cell Ig Gene Transcription, Amplification, and Sequencing

  Construction of complementary DNA (cDNA) from a single B cell provides an efficient method for simultaneous analysis of expressed IgH and IgL genes.  Typically, single B-cell cDNA synthesis is performed in equipment used for cell deposition and cell lysis (96-well plates, nanowell chips, etc.), which ensures easily handling of large numbers of samples and minimizes the risk of cross-contamination.  Full-length Ig variable region gene transcripts were amplified by nested PCR, and RT-PCR products were used as samples for the first round of PCR for further reactions.

  Regardless of the Ig variable region gene amplification strategy used, the transcriptional information of the Ig variable region genes encoding antibody-specific single B-cells was subsequently sequenced.  Then using various databases such as NCBI's IgBLAST and IMGT, the rearranged V, D and J gene segments can be easily identified and analyzed for mutations, insertions and deletions.

• Case Study: Nectin-4 Mouse Serum Titer

  

  Case Study: Nectin-4 (4-1) Sorting Single B Cell

  

  Case Study: Nectin-4 (4-1) Sequencing (Top20)

  

  Case Study: Nectin-4 (4-6) Sorting Single B Cell

  

  Case Study: Nectin-4 (4-6) Sequencing (Top20)

  

FAQ

• Preparation of Monoclonal Antibodies

  (1) Hybridoma technology, including chimeric antibody preparation technology;

  (2) Screen library including phage antibody library technology and ribosome display technology;

  (3) Genetic mice preparation antibody technology

  (4) Single-cell screening technology;

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