Characterization of a human telomerase reverse transcriptase sequence containing two antigenic epitopes with high...

Area of Science:

• Oncology
• Immunology
• Molecular Biology

Background:

• Telomerase activity is elevated in most human malignant tumors but not in normal adult tissues.
• Human telomerase reverse transcriptase (hTERT) is a critical component of telomerase and a potential cancer biomarker.

Purpose of the Study:

• To identify and characterize tumor-associated antigen (TAA) peptides derived from hTERT for cancer diagnosis and prognosis.
• To generate polyclonal antibodies against a specific hTERT-derived TAA for detecting telomerase activity.

Main Methods:

• Selected an hTERT peptide sequence with high affinity for HLA-A2 using a peptide-motif scoring system.
• Cloned the hTERT sequence into an expression vector and expressed it in E. coli.
• Purified the fusion protein, immunized mice to generate anti-hTERT antibodies, and validated antibody specificity using Western blotting on the THP-1 cell line.

Main Results:

• Successfully expressed and purified a GST-hTERT fusion protein.
• Generated polyclonal antibodies that specifically detected hTERT expression in the THP-1 leukemia cell line.
• The identified hTERT antigenic peptides show potential as universal markers for tumor diagnosis and prognosis.

Conclusions:

• The study demonstrates the feasibility of using specific hTERT antigenic peptides as biomarkers for cancer.
• The generated anti-hTERT antibodies can detect telomerase activity, supporting their application in cancer diagnostics and prognostics.