Strategies Insuring the Optimal use of IgG or FAB* Fragments Covalently Bound to 1.4 NM Nanogold ™ in Immunogold Labeling Procedures

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Gold particles as markers in immunocytochemical procedures have undergone tremendous changes since their introduction by Faulk and Taylor (1). With the development of both small colloidal gold (<3 nm) and 1.4 nm gold clusters (Nanogold™) for immunocytochemical labeling, steric hindrance has been significantly reduced. In a comparative study using colloidal gold-anti CD3 complexes and particle sizes of 5 nm, 3 nm and less than 3 nm on surface labeling of lymphocytes there was a 1.5 fold increase in labeling density when 3 nm particles were used. There was no increase when smaller particles were used (unpublished data). In another unpublished experiment, labeling densities on an astrocytoma cell line were compared between 3nm colloidal gold-CD44 Fab’ fragments and 1.4 nm gold-CD44 Fab’ fragments. Again there was no detectable differences between particle sizes. The limiting factors remain the size of the IgG or Fab’ fragment, the ability to bind it whether by absorption or by covalent bonding without rendering it inactive and also the accessibility of the target molecule in the tissue.