Monoclonal antibodies against human telomerase reverse transcriptase (hTERT): preparation, characterization, and application.
The catalytic subunit of human telomerase reverse transcriptase (hTERT), is the necessary and rate-limiting component to telomerase activation in cancer cells. To develop monoclonal antibodies (MAbs) against hTERT, a peptide-hTERT(9)-derived from specific motif T of hTERT was synthesized. Through fusion of splenocytes of BALB/c mice immunized with hTERT(9) with mouse myeloma cells, hybridomas were generated and clones secreting anti-hTERT(9) antibody were screened. After three rounds of limited dilution of candidate clones, three of which present stable and constant antibody production. The MAbs were hTERT(9)-specific and reactive with native hTERT of human cancer cells or tissues in Western blot and immunohistochemistry. The heavy chain variable regions from three hybridomas were cloned and sequenced confirming their mouse Ig derivation. The described investigation suggested that the generated MAbs to hTERT(9) could recognize native hTERT and be useful to cancer research.
Yang
S
S
Department of Pathology, The Health Science Center of Peking University, Beijing 100083, China.
Zhang
B
B
Wang
J
J
Liao
S
S
Han
J
J
Wei
J
J
Hou
L
L
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis*; D000917:Antibody Formation; D015153:Blotting, Western; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006367:HeLa Cells; D006801:Humans; D007124:Immunoenzyme Techniques; D007135:Immunoglobulin Variable Region / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000276:immunology; D010446:Peptide Fragments / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D019098:Telomerase / Q000276:immunology*; D014407:Tumor Cells, Cultured / Q000502:physiology
D000911:Antibodies, Monoclonal; D004268:DNA-Binding Proteins; D007135:Immunoglobulin Variable Region; D010446:Peptide Fragments; D019098:Telomerase