Radioimmunoassay determinations of murine β2‐microglobulin and an H‐2 antigen‐like serum component

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Abstract
Normal mouse serum contains H‐2 antigen‐like components. One such protein consists of a 40000 dalton chain and β2‐microglobulin (β2m) and the other component contains the two H‐2 antigen‐like subunits and, in addition, two larger polypeptide chains with apparent molecular weights (mol. wt.) of 65000 and 80000. Radioimmunoassay procedures specific for the H‐2 antigen‐like chain and β2m, respectively, have been developed. Whereas free β2m and β2m associated with other polypeptide chains reacted identically in the β2m radioimmunoassay, the two serum forms of the H‐2 antigen‐like chain displayed different reactivities in the heavy‐chain assay. The low mol. wt. protein complex, comprising the 40000 dalton chain and β2m, inhibited in the heavy‐chain assay identically with the standard. However, the high mol. wt. complex containing the H‐2 antigen‐like chain gave rise to a biphasic inhibition profile. This was apparently due to some antigenic determinants of the 40000 dalton chain being buried by the 65000 and 80000 dalton chains. Inhibition curves parallel with the standard were, however, obtained on dissociation of the high mol. wt. protein complex with sodium dodecyl sulfate. Thus, by subjecting serum to the ionic detergent‐sensitive radioimmunoassay, procedures for the two H‐2 antigen‐like chains could be developed. The serum levels of the H‐2 antigen‐like chain and of β2m vary with the age of the animal, and adult levels are recorded only at 6 to 7 weeks of age. The serum concentrations of the two proteins are strongly correlated. The H‐2 haplotype appears to partly regulate the serum levels of the two proteins. Mice of the H‐2f haplotype contain about 40‐fold lower levels of the H‐2 antigen‐like chain than do mice of other strains.