Human interleukin-1 receptor antagonist: large-scale expression in Bacillus brevis 47-5Q.
Interleukin-1 receptor antagonist (IL-1RA) has been used as a tool to study the biologic activity of IL-1 and as a possible therapeutic substance for inflammatory disease. To perform in vivo study, however, large quantities of IL-1RA are required. Bacillus brevis strains secrete large amounts of protein but little protease into the medium. Using B. brevis 47-5Q, we developed a large-scale expression system of human IL-1RA (HuIL-1RA). The bacteria secreted HuIL-1RA into the culture medium at very high levels, approximately 200 mg/L. The protein was isolated in one-step purification with monoclonal antibody (mAb) against HuIL-1RA. The IL-1RA molecule was determined to be functionally active by the inhibiting assay of HuIL-1-induced cell proliferation in a mouse T cell line, D10N4M.
Takii
T
T
Department of Hygienic Chemistry, Faculty of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan.
Honda
H
H
Sasayama
S
S
Kobayashi
T
T
Ikezawa
H
H
Udaka
S
S
Oomoto
Y
Y
Onozaki
K
K
D000818:Animals; D001407:Bacillus; D001483:Base Sequence; D004815:Epidermal Growth Factor / Q000096:biosynthesis; D005820:Genetic Testing; D005822:Genetic Vectors; D019382:Human Growth Hormone / Q000096:biosynthesis; D006801:Humans; D053590:Interleukin 1 Receptor Antagonist Protein; D007376:Interleukin-2 / Q000096:biosynthesis; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D012795:Sialoglycoproteins / Q000096:biosynthesis*; D013601:T-Lymphocytes / Q000378:metabolism; D014170:Transformation, Genetic
C507763:IL1RN protein, human; C507764:Il1rn protein, mouse; D053590:Interleukin 1 Receptor Antagonist Protein; D007376:Interleukin-2; D011994:Recombinant Proteins; D012795:Sialoglycoproteins; D019382:Human Growth Hormone; D004815:Epidermal Growth Factor