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Development of Monoclonal Antibodies Against a Riboflavin‐Tryptophan Photoinduced Adduct: Reactivity to Eye Lens Proteins *
Author(s) -
Diaz Marisol,
Becker M. Ines,
Ioannes Alfredo E.,
Silva Eduardo
Publication year - 1996
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.1996.tb09628.x
Subject(s) - tryptophan , riboflavin , adduct , reactivity (psychology) , monoclonal antibody , lens (geology) , chemistry , biochemistry , antibody , biology , genetics , medicine , amino acid , organic chemistry , paleontology , alternative medicine , pathology
We describe here the development of monoclonal antibodies to the hapten tryptophan‐riboflavin, generated by irradiation of a solution of bovine serum albumin in the presence of riboflavin. The specificity of the three obtained monoclonal antibodies, named lE6, 5H5, 5AS all belonging to the IgGl isotype, was assessed by a competitive enzyme‐linked immunosorbent assay in the presence of an increasing concentration of the tryptophan‐riboflavin adduct, obtained from an irradiated riboflavin‐sensitized tryptophan solution. It was demonstrated that the tryptophan‐riboflavin antibodies react with the soluble proteins of the eye lens; this reaction was more intense in the old rat lenses as compared to the young ones, and a maximum binding of the antibodies was obtained with the soluble protein fraction from the human catar‐actous lens. By indirect immunofluorescence, a reactivity associated with the protein matrix, localized in the lens central zone, was observed. In the peripheral zone of the lens, where the younger cells are found, a marked im‐munofluorescent emission was observed on structures preferentially localized in the nuclei.