The invention relates to an AFP recombinant protein, and an in vitro high-efficiency recombination expression method, and specifically relates to a recombinant protein of artificial signal peptide-AFP-609aa-6his, and an in vitro high-efficiency recombination expression method of mammal transient expression system. DNA sequence of the AFP recombinant protein is represented by SEQ ID No.1. The in vitro high-efficiency recombination expression method comprises following steps: a, DNA sequence of the AFP recombinant protein is optimized; b, gene of the AFP recombinant protein is cloned into pcDNA3.1+ carrier via double enzyme digestion, pcDNA3.1+-AFP expression plasmid is constructed, and plasmid extraction is performed; c, transfection of pcDNA3.1+-AFP plasmid, which is obtained via extraction, into CHO-S suspension cells is realized using PEI, and recombinant protein expression is preformed; and d, 7 days later, the AFP recombinant protein is obtained via separation and purification. Activity of the AFP recombinant protein is close to that of natural AFP protein, wherein the artificially synthesized AFP recombinant protein with 6HIS labels is prepared via large-scaled transient suspension CHO-S expression system; purity is relatively high; endotoxin content is low; no pollution is caused by animal-based protein; and it is possible for the AFP recombinant protein to be used as a tumor antigen vaccine in liver cancer immunotherapy.