Preparation of mannan-protein conjugates using high-temperature glycation.

In the present paper, we describe a procedure for the preparation of mannan-protein conjugates. Mannan from Saccharomyces cerevisiae (baker's yeast) was oxidized by NaIO4 and coupled with BSA by means of high-temperature glycation. Depending on the temperature and degree of mannan oxidation, the procedure enabled different levels of substitution to be obtained. For conjugate prepared under selected conditions, the average substitution level was 1.64 molecules of mannan per albumin molecule. The mannan-albumin conjugate was purified by means of ion-exchange chromatography on CM-cellulose at a pH below the pI of albumin. The influence of coupling conditions on protein activity was examined using urease as an example. Urease linked to mannan retained its activity. The mannan-protein conjugates can be used, for example, as carriers of therapeutic substances to macrophages.