The application relates to the field of biological
medicine, and particularly discloses a
monoclonal antibody combined with human CD8 and a preparation method and application thereof. The
antibody is obtained by combining single
B cell sorting,
gene cloning and
mammalian cell expression technology after rabbit immunization of a CD8 polypeptide
antigen, a
New Zealand white rabbit is immunized by adopting a KLH coupled CD8
antigen polypeptide combined with a rapid
immunoadjuvant, high-affinity
antibody clones are obtained by single
B cell sorting technology, and the antibody is prepared by recombination expression. The antibody is verified by ELISA and
flow cytometry, has
high titer and strong specificity, can effectively recognize human
peripheral blood CD8+ T cells, and has a
signal strength and a background
signal-to-
noise ratio which are significantly better than those of a traditional mouse-derived antibody. The application provides light and
heavy chain variable region
amino acid sequences of the antibody, realizes controllable preparation of the antibody and protection of
intellectual property rights. The antibody can be widely applied to the fields of
flow cytometry detection,
immunohistochemistry, tumor
immunotherapy monitoring and
T cell function research, and has important scientific research and clinical application values.