A method for screening positive single B cells that secrete target substances and a method for preparing monoclonal antibodies.

Abstract

This invention relates to the field of biotechnology and provides a method for screening positive single B cells that secrete target substances and a method for preparing monoclonal antibodies. The positive single B cell screening method includes the following steps: isolating PBMCs from the peripheral blood of immunized animals; sorting memory B cells using magnetic beads and separation antibodies; activating and culturing memory B cells in vitro using an activation medium; and loading, analyzing, and exporting target positive cells from the activated memory B cells using a Beacon single-cell photoconductive system. This invention achieves high-throughput screening of FYL positive single B cells through the Beacon single-cell photoconductive system. Gene products are obtained through reverse transcription and PCR amplification, and accurate sequences are obtained through sequence analysis. The synthesized plasmid is expressed in a eukaryotic system, and high-purity FYL rabbit monoclonal antibodies are obtained through affinity chromatography, laying the foundation for the development of a rapid fentanyl detection kit.

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