Construction of human single-chain variable fragment antibodies of medullary thyroid carcinoma and single photon emission computed tomography/computed tomography imaging in tumor-bearing nude mice.
Medullary thyroid carcinoma (MTC) is a rare tumor of the endocrine system with poor prognosis as it exhibits high resistance against conventional therapy. Recent studies have shown that monoclonal antibodies labeled with radionuclide have become important agents for diagnosing tumors. To elucidate whether single-chain fragment of variable (scFv) antibody labeled with 131I isotope is a potential imaging agent for diagnosing MTC. A human scFv antibody library of MTC using phage display technique was constructed with a capacity of 3x10(5). The library was panned with thyroid epithelial cell lines and MTC cell lines (TT). Western blotting and enzyme-linked immunosorbent assay (ELISA) were used to identify the biological characteristics of the panned scFv. Methyl thiazolyl tetrazolium (MTT) assay was also used to explore the optimal concentration of the TT cell proliferation inhibition rate. They were categorized into TT, SW480 and control groups using phosphate-buffered saline. Western blotting showed that molecular weight of scFv was 28 kDa, cell ELISA showed that the absorbance of TT cell group was significantly increased (P=0.000??) vs. the other three groups, and MTT assay showed that the inhibition rate between the two cell lines was statistically significantly different (P<0.05) when the concentration of scFv was 0.1, 1 and 10 µmol/l. The tumor uptake of 131I-scFv was visible at 12 h and clear image was obtained at 48 h using the single photon emission computed tomography. scFv rapidly and specifically target MTC cells, suggesting the potential of this antibody as an imaging agent for diagnosing MTC.
Department of Nuclear Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, P.R. China.
Qiong
Q
Liu
Department of Nuclear Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, P.R. China.
Hua
H
Pang
Department of Radiology, The First Affiliated Hospital of Luzhou Medical College, Chongqing, P.R. China.
Xiaoli
X
Hu
Department of Nuclear Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, P.R. China.
Wenbo
W
Li
Department of Nuclear Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, P.R. China.
Jimei
J
Xi
Department of Nuclear Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, P.R. China.
Lu
L
Xu
Department of Nuclear Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, P.R. China.
Jing
J
Zhou
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D018278:Carcinoma, Neuroendocrine / Q000175:diagnosis* / Q000276:immunology; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D006801:Humans; D007457:Iodine Radioisotopes / Q000378:metabolism*; D008297:Male; D051379:Mice; D009368:Neoplasm Transplantation; D019151:Peptide Library; D057127:Single-Chain Antibodies / Q000276:immunology*; D013964:Thyroid Neoplasms / Q000175:diagnosis* / Q000276:immunology; D015899:Tomography, Emission-Computed, Single-Photon / Q000379:methods*
D000911:Antibodies, Monoclonal; D007457:Iodine Radioisotopes; D019151:Peptide Library; D057127:Single-Chain Antibodies