Construction and expression of antibody-tumor necrosis factor fusion proteins.
The construction, expression and secretion of two genetically engineered antibody-cytokine hybrid fusion proteins is described. To target tumor necrosis factor (TNF) to tumor cells, recombinant antibody techniques were used to generate F(ab')2-like antibody-TNF fusion proteins. At the gene level, an antitransferrin receptor antibody heavy chain gene was linked to a synthetic gene coding for human TNF. The chimeric heavy chain-TNF genes were introduced into a light chain secreting transfectoma cell line, which was producing the light chain of the same antibody. Cell lines were isolated which secreted antibody-TNF fusion proteins of expected size and composition. Culture supernatant of these cell lines contained TNF cytotoxic activity towards murine L929 cells and human MCF-7 cells, indicating that TNF is still active in the fusion protein constructs. These results illustrate the feasibility of the antibody engineering technology to create and produce chimeric mouse-human immunotoxin-like molecules. Furthermore, they demonstrate the ability of mammalian (myeloma) cells to express and secrete antibody-cytokine hybrid molecules with potential use in anticancer therapy.
Hoogenboom
H R
HR
Dr. L. Willems-Instituut, Limburgs Universitair Centrum, Diepenbeek, Belgium.
Volckaert
G
G
Raus
J C
JC
D000230:Adenocarcinoma / Q000188:drug therapy; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D005813:Genes, Synthetic; D006801:Humans; D015151:Immunoblotting; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D066298:In Vitro Techniques; D007951:Leukemia, Myeloid / Q000188:drug therapy; D051379:Mice; D008969:Molecular Sequence Data; D009101:Multiple Myeloma / Q000235:genetics; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000188:drug therapy; D011990:Receptors, Transferrin / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000096:biosynthesis*; D015183:Restriction Mapping; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics* / Q000276:immunology
D007140:Immunoglobulin Fab Fragments; D007143:Immunoglobulin Heavy Chains; D011990:Receptors, Transferrin; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha