Objective By using specific c7c TNF-α mimotope displayed on phage as a target, we got specific TNF-α binding-peptides clones from 12mer phage display peptide library and developed a novel approach for screening phage peptide library. Methods Using c7c TNF-α mimotope phage clones LCS-7 (c-RRPAQSG-c) as a target to screen 12mer phage peptide library and identify the positive clones by phage ELISA and competive ELISA. Results After 3 rounds of screening, 10 of 20 clones were identified as positive clones and all the 10 phage clones shared the identical amino acid sequence: EHMALTYPFRPP. Conclusion The positive 12mer phage clone peptide screened with cycle 7mer TNF-α simulating phage clone as a target can bind to TNF-α specifically. The results suggest that this method is very specific and easy- for screening of binding epitope from phage display peptide library. [