Blocking of non-specific sorption in ELISA on formylated polystyrene beads.
The effect of pH of the blocking solution on the non-specific sorption of peroxidase conjugates to the 0.63-cm formylated polystyrene beads was studied. Anti-G-HRP and BSA-HRP sorption was shown to dramatically depend on pH of blocking protein solutions and was minimum at pH=pI. Sorption of G-HRP weakly depended on pH of the blocking buffer. The blocking efficiency of proteins on unmodified beads, as well as of Tween-20 on formylated beads appeared to be pH independent. The optimized blocking step resulted in a multiple decrease in the non-specific sorption of conjugates on formylated beads.
Shmanai
V V
VV
Institute of Physical Organic Chemistry, Belarussian Academy of Sciences, Minsk. ifoch@ifoch.bas-net.by
D000327:Adsorption; D000818:Animals; D000888:Antibodies, Anti-Idiotypic; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005069:Evaluation Studies as Topic; D006735:Horseradish Peroxidase; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G; D010316:Particle Size; D011137:Polystyrenes; D012710:Serum Albumin, Bovine; D012756:Sheep
D000888:Antibodies, Anti-Idiotypic; D007074:Immunoglobulin G; D011137:Polystyrenes; D012710:Serum Albumin, Bovine; D006735:Horseradish Peroxidase