Immobilization of Fv antibody fragments on porous silica and their utility in affinity chromatography.

Recent advances in molecular biology have allowed antibody binding domains to be cloned and expressed in Escherichia coli. The use of Fv antibody fragments as ligands in immunoaffinity chromatography is reported. Fv fragments specific for hen-egg lysozyme were immobilized on porous silica and used to recover antigen from spiked serum in a single step. Comparison with a conventional immunoadsorbent (whole antibodies immobilized on silica) showed the Fv-silica to have a fivefold superior capacity. Analysis of sectioned Fv-silica particles by immunoelectron microscopy indicated that captured antigen was evenly distributed throughout the internal porous structure of the particle.