Although patients with drug-resistant temporal lobe epilepsy (TLE) achieve good seizure control with resective surgery combined with anti-epileptic drugs (AEDs) treatment, it is still not clear when to withdraw AEDs after the surgery due to a lack of objective strategy for assessing the surgery outcome. Our aim is to identify objective and efficient biomarkers for assessment of surgery outcome to guide postoperative pharmacotherapy. Here, we detected 469 proteins differentially expressed between pre- and post-operative sera of TLE patients with postoperative seizure freedom, and identified six candidate proteins (HBA1, HBB, PRDX2, CA1, CAT and BLVRB) for validation. These six proteins were significantly reduced within three months after surgery, indicating these proteins may be potential biomarkers to assess the surgery outcome for TLE patients in the early stage. Interestingly,we found HBA1 to HBB ratio was superior to individual proteins to evaluate the postoperative surgery outcome with 90.0% sensitivity, 90% specificity and area under the curve of 0.960. We also suppose these proteins were derived from epilepsy foci due to the rat experiment results that HBA1, HBB, PRDX2, CA1 and CAT were highly expressed in cerebrospinal fluid of epilepsy rat models. Additionally, HBA1, HBB, PRDX2, CA1 and BLVRB proteins were highly expressed in the sera of TLE patients with postoperative seizure freedom when compared to TLE patients with postoperative seizure and patients with other types of epilepsy, suggesting surgery outcome or the original sites of seizure might be assessed before the operation through monitoring these protein levels in the sera. Our study provides experimental data for establishing objective indicators for assessing seizure freedom after resective surgery and also proposed potential markers for specific diagnoses of TLE and prediction of TLE surgery outcome.