Simple enzyme immunoassay methods for recombinant human tumor necrosis factor alpha and its antibodies using a bacterial cell wall carrier.
We have developed simple methods for measuring recombinant human tumor necrosis factor alpha (rHu-TNF alpha) and antibodies to rHu-TNF alpha in the sera of animals intravenously injected with rHu-TNF alpha. rHu-TNF alpha was measured by a competitive binding enzyme immunoassay (C-EIA) using standard rHu-TNF alpha, beta-galactosidase labeled rHu-TNF alpha as enzyme-labeled antigen (E-Ag) and anti-rabbit IgG goat immunoglobulins coupled to bacterial cell walls (insolubilized second antibody). In contrast, anti-rHu-TNF alpha antibodies were measured by a sandwich EIA (S-EIA) using purified anti-rHu-TNF alpha rabbit IgG as standard, beta-galactosidase labeled rHu-TNF alpha as E-Ag, and rHu-TNF alpha coupled to bacterial cell walls as insolubilized antigen. C-EIA permits the determination of serum rHu-TNF alpha within the range of 2-150 U/ml (about 0.7-52 ng/ml) with a CV of below 7.6% and 99% recovery. S-EIA permits the determination of anti-rHu-TNF alpha antibodies within the range of 70-1000 ng/ml with a CV of less than 4% and 94.8-106.9% recovery.
Sunahara
N
N
Research Laboratories, Dainippon Pharmaceutical Co., Ltd., Osaka, Japan.
Kurooka
S
S
Kaibe
K
K
Ohkaru
Y
Y
Nishimura
S
S
Nakano
K
K
Sohmura
Y
Y
Iida
M
M
D000818:Animals; D000906:Antibodies / Q000032:analysis*; D001426:Bacterial Proteins / Q000378:metabolism; D002473:Cell Wall; D004355:Drug Stability; D006207:Half-Life; D006801:Humans; D007124:Immunoenzyme Techniques; D007164:Immunosorbents; D011994:Recombinant Proteins / Q000032:analysis* / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis* / Q000276:immunology
D000906:Antibodies; D001426:Bacterial Proteins; D007164:Immunosorbents; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha