Bispecific antibody-producing hybrid hybridoma and its use in one-step immunoassays for human lymphotoxin.
A hybrid hybridoma cell line secreting a bispecific monoclonal antibody (MAb) was constructed by fusing horseradish peroxidase (HRPO)-immunized mouse splenocytes with previously established mouse hybridomas secreting anti-human lymphotoxin (hLT). This cell line was grown in ascitic fluid in mice to obtain large quantities of hybrid MAbs and a bispecific antibody, reacting with both HRPO and hLT, was separated from the monospecific antibody or other inactive immunoglobulin populations by hydroxylapatite chromatography. Sodium dodecylsulfate-polyacrylamide gel electrophoresis demonstrated that the bispecific antibody molecule contained two different types of heavy and light chains of both anti-HRPO and anti-hLT origin. The bispecific antibody was used to establish one-step enzyme immunoassays (EIAs) employing competitive and sandwich systems. The simple sandwich EIA was able to detect 1-100 U/ml of hLT and there was good correlation (r = 0.96) between hLT concentrations measured by the one-step EIA and a bioassay using L929 cell-lysis.
Tada
H
H
Central Research Division, Takeda Chemical Industries, Ltd., Osaka, Japan.
Toyoda
Y
Y
Iwasa
S
S
D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D002459:Cell Fusion; D002851:Chromatography, High Pressure Liquid; D006735:Horseradish Peroxidase / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D007124:Immunoenzyme Techniques; D008233:Lymphotoxin-alpha / Q000032:analysis*; D009500:Neutralization Tests; D010544:Peroxidases / Q000276:immunology*
D000911:Antibodies, Monoclonal; D008233:Lymphotoxin-alpha; D006735:Horseradish Peroxidase; D010544:Peroxidases