The invention 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 disclosed by the invention comprises VHCDR1, VHCDR2 and VHCDR3, wherein the
amino acid sequences of the VHCDR1, the VHCDR2 and the VHCDR3 are shown as SEQ ID NO: 1-3; the
amino acid sequences of the VLCDR1, the VLCDR2 and the VLCDR3 are as shown in SEQ ID NO: 4-6. The
antibody development process comprises the following steps: synthesizing Human CCR8mRNA, immunizing a mouse, collecting
spleen,
bone marrow and
lymph node cells of the mouse, wrapping a single
plasma cell suspension, a
fluorescence labeled secondary antibody and a Human CCR8HEK293 overexpression
cell line in water-in-oil liquid drops together by adopting a microfluidic
liquid drop sorting
system, carrying out
dielectrophoresis sorting, and enriching
plasma cells secreting the target
cell binding antibody; carrying out single-cell
library construction on
plasma cells, and carrying out NGS sequencing to obtain antibody sequences with naturally paired light and heavy chains; and selecting an antibody sequence for high-
throughput expression of mammalian cells, and carrying out flow detection
verification. The
monoclonal antibody provided by the invention has good
antigen-antibody binding activity and specificity on the
cellular level.