Effect of agarose variability on the measurement of lysozyme activity

Lysozyme assays are often performed by a diffusion technique utilizing agarose gels impregnated with substrate organisms (lysoplates), but the results differ greatly from those obtained with spectrophotometric or immunologic techniques. We have investigated the effect of agarose composition on the lysoplate assay utilizing 10 different gels varying in ionic parameters. Standard curves generated with purified human lysozyme solutions were parallel, but the diameters of the zones of lysis varied inversely with gel sulfate content. The different agaroses had variable effects on determinations of normal serum lysozyme, and the results obtained on any given gel agreed with neither those found on other gels nor with independent assay in another system. The lysoplate assay should be utilized only in those laboratories that can obtain uniform agarose preparations and extensively calibrate normal ranges for their gels.