The application belongs to the technical field of
molecular biology, and particularly relates to an anti-human CCR8
monoclonal antibody and a preparation method thereof. The anti-human CCR8
monoclonal antibody of the application comprises VHCDR1, VHCDR2 and VHCDR3 with the
amino acid sequences shown in SEQ ID NO: 1-3; and VLCDR1, VLCDR2 and VLCDR3 with the
amino acid sequences shown in SEQ ID NO: 4-6. The
antibody development process is as follows: Human CCR8 mRNA is synthesized, mice are immunized, and
mouse spleen,
bone marrow and
lymph node cells are collected; a microfluidic droplet sorting
system is used to wrap single
plasma cell suspension, fluorescently labeled secondary antibody and Human CCR8 HEK293 overexpression
cell line in water-in-oil droplets, and
dielectrophoresis sorting is performed to enrich
plasma cells secreting target
cell binding antibodies; single cell
library construction is performed on the
plasma cells, and NGS sequencing is performed to obtain light and
heavy chain naturally paired antibody sequences; the antibody sequences are selected for
mammalian cell high-
throughput expression, and
flow cytometry detection
verification is performed. The
monoclonal antibody of the application has good
antigen-antibody binding activity and specificity at the cell level.