The invention discloses a preparation method and application of a cat main
allergen FEL-1 nano
antibody FEL-1-Nb, and relates to the technical field of
antibody preparation, and the preparation method comprises the following steps: preparing an
immunogen; animal immunization and
library construction; constructing and panning a
phage display library; screening and identifying positive clone; and expression and purification of the nano
antibody:
cloning the positively cloned VHH
gene to an
expression vector such as a pET series, transforming an expression strain such as
escherichia coli BL21 (DE3), carrying out IPTG induced expression, and purifying through an inclusion body purification method to obtain a high-purity nano antibody
protein. The nano antibody FEL-1-Nb provided by the invention can be specifically bound with a cat main
allergen FEL-1
protein, the unique
amino acid sequence of the nano antibody FEL-1-Nb is shown as SEQIDNO: 1, high-
affinity binding with the FEL-1
protein is ensured, the positive clone binding rate is high through
phage display panning and ELISA
verification, the phage
recovery rate after the third round of panning is increased by more than 1000 times compared with the first round of panning, and the positive clone binding rate is higher than that of the first round of panning. Therefore, the nano antibody has excellent targeted recognition capability.