Thirty monoclonal antibodies (MoAbs) to human α-fetoprotein (AFP) were compared with one another by two methods: Immunoaffinity electrochromatography or additive ELISA. The first method permitted to analyse the epitopes of native AFP in solution [Abelev et al., Immunol Lett 1994;40:133–138] while the other approach also detects the epitopes of conformationally modified (partly denatured) AFP fixed on the plastic [Yazova et al., Immunol Lett 1990;25:325–330]. Competitive analysis of all MoAbs revealed 10 epitopes, 9 expressed on native AFP and 1 only on the partly denatured molecule. The cross-reactions between separate MoAbs allowed to include them into 6 distinct epitope clusters, or immunodominant groups with the characteristic patterns of reactivity. The obtained epitope map of AFP is necessary for the construction of AFP detection kits as well as for the identification of its antigenic and functional subfractions.
The immunological heterogeneity of human α-fetoprotein (AFP) was demonstrated using immunoaffinity electrochromatography on monoclonal antibodies (MoAbs) to 3 non-cross-reacting epitopes of this protein. At least 4 subfractions expressing different epitopes were found in the native AFP. These subfractions demonstrated molecular weights similar to the major component of the original AFP. The difference between epitope F5-positive and F5-negative subfractions disappeared when epitope-negative subfraction was conformationally changed after fixation onto nitrocellulose membrane (NCM). Thus, the epitope under study exists in two forms on the native human AFP molecule: an open and a cryptic form. The cryptic form could be revealed after partial denaturation by fixation on NCM. The epitope variants of AFP could possess different functions in multifunctional AFP. The AFP epitope heterogeneity found in this work should be taken into account when constructing diagnostic AFP kits and when isolating purified AFP using anti-AFP MoAbs.
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