Preparation and Application of Soluble Tim-3 Recombinant Protein and Mutant Protein Thereof

A purification method of soluble Tim-3 recombinant protein, comprising the following steps:

• performing fluid exchanging on cell culture solutions of eukaryotic CHO cells, prokaryotic E. coli and insect baculovirus expression system containing recombinant human soluble Tim-3 through microfiltration clarification and ultrafiltration concentration, and then passing through cation and anion chromatographic column, molecular sieve chromatographic column, hydrophobic chromatographic column and/or affinity chromatographic column to obtain the soluble Tim-3 recombinant protein with enough purity, wherein the amino acid sequence of the soluble Tim-3 recombinant protein is as shown in SEQ ID NO:
• 1.

A soluble Tim-3 recombinant mutant protein, wherein the mutant protein is a soluble Tim-3 recombinant mutant protein obtained in the mode that:

• on the basis of a gene sequence corresponding to human Tim-3 (NP_116171.3) extracellular domain amino acid (SEQ ID NO:
• 1), mutant genes are obtained by screening through a directed evolutionary technology, and then expressed, wherein the mutant sites of the mutant genes do not involve conserved sequences among species, but occur in non-conservative sequence positions.

A soluble Tim-3 recombinant mutant protein, wherein the amino acid sequence of the soluble Tim-3 recombinant mutant protein is one of the sequences as shown in SEQ ID NOs:

• 2-7.