Characterization of human lens antigens by crossed immunoelectrophoresis

Abstract

Crossed immunoelectrophoresis and fused rocket electrophoresis were used to examine fractions of human lens homogenates obtained by ion exchange (DEAE) and gel (G-200) chromatography. The methods were found useful for identifying the various antigens found in each fraction, to evaluate the purity of the fractions, and to compare both qualitatively and semi-quantitatively, the antigenic content in the various fractions. It is suggested that these immunological techniques may provide the basis for an extended nomenclature of lens crystallins. Considerable differences were found between fractions from lenses of young individuals and from those of old individuals. The fractions from young lenses were fairly homogeneous while those from old lenses were heterogeneous. The DEAE fraction 0·4 m from old human lenses contained, in addition to α-crystallin, considerable amounts of β- and γ-crystallins. It is suggested that this, at least in part, explains the presumed increase in α-crystallin content of human lenses with age. The α-crystallin preparations obtained by ion exchange and gel chromatography were equally pure. The purest β-crystallin preparations were obtained by ion exchange chromatography while the purest γ-crystallin preparation was obtained by gel chromatography.