Antigenic characterization of recombinant, lymphoblastoid, and leukocyte IFN-alpha by monoclonal antibodies.
To gain more insight into similarities of different interferon-alpha (IFN-alpha) species, we evaluated neutralization and immunoactivity of a variety of IFN preparations with various monoclonal antibodies (IFN-alpha mAb). Nine IFN-alpha mAb obtained through immunization with recombinant IFN-alpha (rmAb), lymphoblastoid IFN-alpha (LY mAb), and leukocyte IFN-alpha (LE mAb) were tested. The IFN-alpha mAb were evaluated for their ability to neutralize the antiviral activity of 11 recombinant IFN-alpha subtypes, two recombinant IFN-alpha hybrids, and lymphoblastoid and leukocyte IFN-alpha preparations. The same IFN-alpha mAb were also used in immunoblotting, and some of them were used in immunoaffinity chromatography. The results of the neutralization assay reveal that the IFN-alpha mAb significantly differ in their ability to neutralize the individual IFN-alpha species. Interestingly, none of the IFN-alpha mAb was able to neutralize all the IFN-alpha species. In particular, rmAb were unable to neutralize LE-IFN-alpha or LY-IFN-alpha, whereas LE mAb and LY mAb efficiently neutralized rIFN-alpha2. In some cases, the epitopes to which IFN-alpha mAb are directed were identified through the use of synthetic fragments of IFN-alpha2 or by evaluating the selectivity in binding to IFN-alpha subtypes.
Viscomi
G C
GC
Biotechnology Department, Alfa Wassermann, Bologna, Italy. GCVISCOMI@ALFAWASSERMANN.IT
Antonelli
G
G
Bruno
C
C
Scapol
L
L
Malavasi
F
F
Funaro
A
A
Simeoni
E
E
Pestka
S
S
De Pisa
F
F
Dianzani
F
F
D000818:Animals; D000911:Antibodies, Monoclonal; D000937:Antigen-Antibody Reactions; D002846:Chromatography, Affinity; D006801:Humans; D015151:Immunoblotting; D007370:Interferon Type I / Q000276:immunology*; D016898:Interferon-alpha / Q000276:immunology*; D007962:Leukocytes / Q000276:immunology*; D008214:Lymphocytes / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D011994:Recombinant Proteins; D013234:Stem Cells / Q000276:immunology*
D000911:Antibodies, Monoclonal; D007370:Interferon Type I; D016898:Interferon-alpha; D011994:Recombinant Proteins