The invention discloses an application of Spy Tag / Spy Catcher in preparation of a
thrombin-
antithrombin III compound, a preparation method of the
thrombin-
antithrombin III compound and an application of the
thrombin-
antithrombin III compound. The preparation method comprises the following steps: respectively obtaining a Spy Catcher-
Thrombin gene segment and a Spy Tag-
Antithrombin gene segment; respectively connecting the
gene segments to starting vectors to form corresponding connectors; carrying out conversion and screening operation on the
linker to obtain a corresponding recombinant
plasmid; carrying out transformation and induction operation on the recombinant
plasmid, so as to obtain a Spy Catcher-
Thrombin recombinant
protein and a Spy Tag-
Antithrombin recombinant
protein; the preparation method comprises the following steps: mixing the Spy Catcher-
Thrombin recombinant
protein with the Spy Tag-
Antithrombin recombinant protein, and carrying out a reaction, so as to obtain the TAT protein. According to the method, the
spontaneous reaction of Spy Tag / Spy Catcher is utilized, other chemical cross-linking agents are not needed, the cost can be reduced,
green production is facilitated, and the recombinant protein can be produced on a large scale by utilizing a
cell culture technology; the recombinant TAT
antigen and the recombinant TAT
antibody prepared by the invention have strong immunoreactivity, and completely meet the requirements of
thrombus detection on the TAT
antigen.