Lysozyme antibody affinity

Determination of Effects of Amino Acid Replacements among Evolutionary Variant Lysozymes on Antibody Affinity... 

Lysozyme has been isolated from bovine and canine cartilages and from canine sera by a variety of chromatographic procedures, including affinity chromatography on carboxymethylchitin. The composition of the bovine lysozyme is similar to that of other mammalian lysozymes. Anti-(canine lysozyme) antibodies cross-reacted with bovine lysozyme, but with neither hen egg-white lysozyme nor embryonic chicken-cartilage lysozyme. Electron microscopy has shown that lysozyme in canine epiphyseal plates is localized extracellularly, mainly in the vicinity of the chondrocytes. Evidence that lysozyme has a physiological role in normal endochondral calcification has been presented. ... 

Lavoie, T. B., Drohan, W. N. and Smith-Gill, S. J. (1992), Experimental analysis by site-directed mutagenesis of somatic mutation effects on affinity and fine specificity in antibodies specific for lysozyme , f. Immunol., 148, 503-513. 

Hawkins RE, Russell SJ, Baier M, Winter G, The contribution of contact and noncontact residues of antibody in the affinity of binding to antigen. The interaction of mutant D1.3 antibodies with lysozyme, J. Mol. Biol., 234 958-964, 1993. 

The objective of the experiments is to compare the affinity of wild-type antibody and wild-type immunogen (chicken lysozyme) with that of wild-type antibody and mutant antigen or with that of mutant antibody and wild-type immunogen. All the assays involve incubation of a constant concentration of one reactant with varying amounts of the complementary reactant, along with estimating the concentration of either bound or free reactant by an immunochemical or enzymatic method. In each case, the assumption is made that the measurement step does not disturb the equilibrium between antigen and antibody, and it is important that this assumption be validated experimentally. We summarize below several alternative methods. 

Fig. 3. Affinity analysis of wild-type and mutant K49VlcT of MAb HyHEHO sequentially saturated with duck lysozyme in the particle affinity assay. The abscissa (R ) is the corrected proportion of antibody binding sites filled with soluble lysozyme, and the ordinate is equivalent to [Ab] times B/F, as described in the text (Reproduced from Lavoie et al.26)...

The conformational energy calculations received support subsequently from two types of experimental study by Smith-Gill et al. (1984). The affinity of ring-necked pheasant lysozyme, in which Asn and Arg at positions 113 and 114 are replaced by Lys and His, respectively, is the same for (NAG)e as that of domestic hen egg-white lysozyme (i.e., the right side is not involved). They also showed that a monoclonal antibody bind-... 

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