The single B-cell antibody technology takes advantage of the fact that a single B cell contains only one functional heavy-chain variable region DNA sequence and one light-chain variable region DNA sequence, and that a single B cell produces only one specific antibody. It involves isolating specific B cells from the tissues or peripheral blood of immunized animals, then using single-cell PCR technology to amplify the heavy-chain and light-chain variable region DNA sequences from B cells secreting a single antibody, and finally expressing them in mammalian cells to obtain biologically active monoclonal antibodies. Currently, the single B-cell antibody technology has been widely applied in the fields of pathogenic microorganism infections, tumors, autoimmune diseases, and organ transplantation, demonstrating unique advantages and promising prospects for future applications.

Antibodies produced using traditional hybridoma technology have a long development cycle, with the immunization process alone typically taking about 8 months. Furthermore, due to limitations in the fusion rate of hybridomas—where only a small fraction of the entire B-cell population undergoes complete fusion—the number of positive clones obtained is low, making this approach unsuitable for comprehensive screening of large antibody libraries. As for phage display technology, although it offers a large library capacity, the pairing of the heavy chain variable region (VH) and light chain variable region (VL) typically relies on random combinations, resulting in mostly unnatural VH-VL antibody pairings. In contrast, single B-cell antibody technology—as a next-generation antibody development technique following hybridoma and phage display technologies—offers advantages such as a shorter development cycle, a higher number of positive clones, fully human antibodies, and a reduced requirement for cell numbers. Most importantly, antibodies obtained through single B-cell antibody technology ensure a natural pairing of light and heavy chains, which is crucial for antibody development.

The entire process of single B-cell antibody technology consists of five main steps: B-cell enrichment, single B-cell screening, antibody gene sequencing and analysis, recombinant antibody expression, and antibody function validation.