Radioiodination of human interferon-alpha2 interferes with binding of C-terminal specific antibodies.
Radioimmunoassays based on reactivity between a monoclonal antibody (mAb) and human 125I-interferon (IFN)-alpha2 are frequently exploited in interferon research. In general, epitopes of antibodies specific for human IFN-alpha2 are located on the two immunodominant structures formed in the N- and C-terminal domains, respectively. We found that labelling of IFN-alpha2 with Na(125)I by the chloramine-T method did not affect the binding of antibodies recognising the N-terminal region 30-53. In contrast, radioiodination of IFN was associated with a dramatic decrease in IFN reactivity with mAbs specific for the C-terminus (residues approximately 120-145 approximately ). We suggest that steric hindrance araising from the incorporation of 125I into the tyrosine residues at positions 123, 130 and 136 may be responsible for the change in immunoreactivity. The adverse effect of radioiodination of IFN-alpha2 on the binding potency of C-terminal specific mAbs must be taken into consideration in experiments based on the interaction of such antibodies (i.e. NK2) with the labelled antigen.
Schmeisser
H
H
Institute of Virology, Slovak Academy of Sciences, 842 46, Bratislava, Slovakia.
Kontseková
E
E
Vancová
I
I
Mucha
V
V
Kontsek
P
P
D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D000937:Antigen-Antibody Reactions; D006801:Humans; D016898:Interferon-alpha / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D007457:Iodine Radioisotopes; D007553:Isotope Labeling / Q000379:methods; D011487:Protein Conformation; D011863:Radioimmunoassay; D011994:Recombinant Proteins
D000911:Antibodies, Monoclonal; D016898:Interferon-alpha; D007457:Iodine Radioisotopes; D011994:Recombinant Proteins; C554129:interferon alfa-2c