The invention discloses a pig whole
sperm membrane protein antigen specific nano
antibody library construction method which comprises the following steps: S1, immunizing alpaca with pig whole sperms, and separating
peripheral blood cells after immunization; s2,
RNA in
peripheral blood cells is extracted, cDNA is obtained through reverse transcription, and a VHH coding sequence of an
antibody is amplified; s3,
cloning the amplified VHH coding sequence to a
bacteriophage plasmid vector to prepare a recombinant
bacteriophage vector; s4, transforming the recombinant
bacteriophage vector into
escherichia coli and carrying out enlarged culture to obtain a bacteriophage display
library; and S5, performing panning on the
phage display library for a plurality of times by using the pig X / Y sperms, and constructing the pig single sex
sperm specific nano
antibody library. According to the method, the pig whole
sperm is adopted as an
immunogen for immunizing alpaca for the first time, the capacity of the constructed antibody library reaches 3.4 * 10 < 8 > pfu, the abundance of the antibody library is higher, the coverage degree of whole sperm antigens is higher, and the variety is more complete, so that the capture success rate of the pig sex control
specific antibody is increased, and a foundation is laid for industrial application of the pig sperm
specific antibody.