Specific recognition of cytosolic thymidine kinase in the human lung tumor by monoclonal antibodies raised against recombinant human thymidine kinase.
Anti-TK monoclonal antibodies (mAbs) were raised against recombinant human cytosolic thymidine kinase (rhTK) and characterized by Western immunoblotting, enzyme-linked immunosorbent assay (ELISA) and immunostaining of tumor cells. Twenty-three clones of TK mAbs were characterized to recognize specifically not only rhTK produced by Escherichia coli but also TK subunit of 25 kDa in human lung cancer. The anti-TK mAbs reacted specifically with cytosolic TK but not with mitochondrial TK. Only one clone of the mAbs inhibited the catalytic activity of TK. By solid phase sandwich enzyme immunoassay using these mAbs, we could quantitate the cytosolic TK content in tissues. Immunohistochemical staining analysis using one of the TK mAbs showed that human lung adenocarcinoma and squamous cell carcinoma exhibited much higher staining intensity than stromal cells. These mAbs are useful for biochemical studies on the regulation of human TK in proliferating cells such as tumor cells and for diagnosis of highly proliferating tumors.
Kuroiwa
N
N
Department of Biochemistry, Chiba University School of Medicine, 1-8-1, Inohana, Chuo-ku, 260-8670, Chiba, Japan. nkuroiwa@med.m.chiba.u-ac.jp
Nakayama
M
M
Fukuda
T
T
Fukui
H
H
Ohwada
H
H
Hiwasa
T
T
Fujimura
S
S
D000230:Adenocarcinoma / Q000175:diagnosis / Q000201:enzymology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D014408:Biomarkers, Tumor / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D015153:Blotting, Western; D002294:Carcinoma, Squamous Cell / Q000175:diagnosis / Q000201:enzymology; D002457:Cell Extracts / Q000032:analysis; D003600:Cytosol / Q000201:enzymology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D007150:Immunohistochemistry; D008175:Lung Neoplasms / Q000175:diagnosis* / Q000201:enzymology; D051379:Mice; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology; D013937:Thymidine Kinase / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; D002457:Cell Extracts; D021122:Protein Subunits; D011994:Recombinant Proteins; D013937:Thymidine Kinase