Monoclonal antibodies specific to chicken iodopsin

Monoclonal antibodies (mABs) from hybridoma cells were raised and screened with a purified cone pigment, iodopsin, from the chicken retina. Four different methods were used to test these antibodies: (1) dot-immunobinding assay; (2) enzyme-linked immunoabsorbent assay (ELISA); (3) one dimensional immunoblotting and (4) two dimensional immunoblotting. Three classes of antibody producing cell lines were identified. One class produces a mAB specific to iodopsin. The mAB from the second class crossreacts with iodopsin and probably one of the other three cone pigments. The mAB from the third class probably crossreacts with all the four cone pigments in the chicken retina. The mABs from all these classes of hybridoma cell lines were selected so that they do not crossreact with rhodopsin. Two dimensional immunoblotting indicated that iodopsin has a much higher isoelectric point than rhodopsin, as suggested from the known amino acid sequences of human rod and cone pigments.