Establishment of antihuman IFN-alpha8-specific monoclonal antibodies and their application in the enzyme-linked immunosorbent assay (ELISA).
In the present study, we describe the generation of a series of anti-interferon-alpha8 (IFN-alpha8)-specific monoclonal antibodies (mAbs), their characterization, and the establishment of a sandwich enzyme-linked immunosorbent assay (ELISA) system for human IFN-alpha8. The sandwich ELISA system is highly sensitive to human natural IFN-alpha8 (nIFN-alpha8), with a minimum detection limit of 50 pg/mL, which did not cross-react with the other IFN preparations and several cytokines tested. Using this ELISA system, pharmacokinetic properties of an IFN-alpha preparation administered in mice were examined. We found that IFN-alpha8 has higher vascular permeability and stability than IFN-alpha2 in the circulation. These results suggest that this ELISA would be very useful for determination of IFN-alpha8 protein concentrations in various experimental samples and also of pharmacokinetic properties of IFN-alpha preparations in human.
Ushio
Chie
C
Biomedical Institute, Research Center, Hayashibara Biochemical Laboratories, Inc., Okayama, Japan.
Ariyasu
Harumi
H
Kayano
Tohru
T
Ohta
Hitomi
H
Aga
Miho
M
Ariyasu
Toshio
T
Ohta
Tsunetaka
T
Kurimoto
Masashi
M
Fukuda
Shigeharu
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification / Q000493:pharmacokinetics; D002199:Capillary Permeability; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D005260:Female; D006801:Humans; D006825:Hybridomas; D016898:Interferon-alpha; D007372:Interferons / Q000276:immunology* / Q000493:pharmacokinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D051381:Rats; D011914:Rats, Inbred BN; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D000939:Epitopes; C554164:IFNA8 protein, human; D016898:Interferon-alpha; D007372:Interferons