Production of a protein A--lymphotoxin hybrid protein for cancer-targeted therapy.
A hybrid protein between staphylococcal protein A and human lymphotoxin, ALT, was produced in Escherichia coli by expression of a recombinant plasmid containing the respective genes. IgG-binding activity of ALT was confirmed by Western blotting analysis and by affinity purification with IgG-Sepharose column chromatography. The purified ALT had cytotoxicity on mouse L-929 cells and its specific activity was approximately 3.5-5.0 X 10(6) U mg-1. ALT was partially degraded by a protease including in the E. coli lysate or trypsin and was converted to lymphotoxin which lacks the NH2-terminal 19 residues but possesses higher cytotoxic activity than ALT.
Genetic Engineering Section, Sagami Chemical Research Center, Kanagawa, Japan.
M
M
Wada
T
T
Kasumimoto
T
T
Miki
H
H
Osada
S
S
Kato
D000595:Amino Acid Sequence; D001483:Base Sequence; D001709:Biotechnology; D004247:DNA / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D000922:Immunotoxins / Q000096:biosynthesis* / Q000235:genetics; D008233:Lymphotoxin-alpha / Q000096:biosynthesis* / Q000235:genetics; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D013205:Staphylococcal Protein A / Q000096:biosynthesis* / Q000235:genetics; D014357:Trypsin
D000922:Immunotoxins; D008233:Lymphotoxin-alpha; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D004247:DNA; D014357:Trypsin