Application of Namalva interferon-alpha monoclonal antibodies for purification and enzyme immunoassay of interferon-alpha.
Namalva (or Namalwa) interferon (IFN)-alpha was partially purified using a combination of conventional methods and modified acid-ethanol extraction. Four mouse monoclonal antibodies against Namalva IFN-alpha were prepared by hybridoma technology after immunization with Namalva IFN-alpha thus purified. Three of these monoclonal antibodies recognized the same or a similar epitope on Namalva IFN-alpha. One of these antibodies was paired with the fourth recognizing a different epitope and used respectively as enzyme-conjugated antibody and solid-phase antibody in our one step enzyme immunoassay (EIA) for IFN-alpha. This assay is simple and was able to detect as little as 5 pg of IFN-alpha in 100 microliters of sample in the short time of 5 hr. There was a good correlation between the EIA and bioassay. The use of one of the monoclonal antibodies as an immunoadsorbant to purify Namalva IFN-alpha is also described.
Notani
J
J
Research Laboratories, Fujisawa Pharmaceutical, Co., Ltd., Osaka, Japan.
Kaizu
T
T
Mutoh
S
S
Otsuka
K
K
Kusunoki
C
C
Yamada
H
H
Nagayoshi
A
A
Suzuki
S
S
Niwa
M
M
Ueda
I
I
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000918:Antibody Specificity; D002460:Cell Line; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D006801:Humans; D006825:Hybridomas; D007124:Immunoenzyme Techniques; D007370:Interferon Type I / Q000032:analysis*; D007962:Leukocytes / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007370:Interferon Type I