Abstract: To screen peptides with high affinity to cryptotanshinone, using cryptotanshinone as a ligand, phage clones binding to cryptotanshinone were screened with phage display 12-mer peptide library through three rounds of biopanning. After binding activity was measured by ELISA, the phage clones displaying high affinity to cryptotanshinone were selected and amplified. DNAs of phage clones were extracted and sequenced, and the coding peptide sequences were analyzed by bioinformatics. Amino acid sequences of 10 positive clones with high affinity to cryptotanshinone were identified. 614 proteins matched partially to the peptide found by homological retrieval with BLAST. The core peptide sequence VILDFGEI was deduced by this study. Peptides with high affinity to cryptotanshinone were obtained through the experiment, which provided experimental evidence and structural foundations for further study of the targets of effects and molecular mechanisms of actions of cryptotanshinone.