Abstract
beta-(1,6)-Branched beta-(1,3)-D-glucans like schizophyllan from the basidiomycete Schizophyllum commune excite various immunostimulatory effects and have been clinically tested as adjuvants. Some of the glucans are also applicable in food or petrol industry due to their viscosity and temperature stability in aqueous solution. Antibodies against these glucans could be used as tool for analysis of glucan preparations or for further research of its bioactivity. Therefore, an immune phage display library was constructed from mice immunized with schizophyllan. Three recombinant monoclonal antibodies were isolated from this library by affinity selection (panning) on schizophyllan. The half-maximal effective concentration (EC50) values for those antibodies varied between 16.4 ng mL−1 and 21.3 ng mL−1. The clones showed binding specificity not only for schizophyllan but also for other beta-(1,6)-branched beta-(1,3)-D-glucans of similar macromolecular structure. Denaturation of the secondary structure led to a reduced antibody binding, indicating an epitope requiring the correct conformation of the triple helical structure of the glucans.
Highlights
A variety of fungal beta-(1,3)-D-glucans are described as substances with antitumoral and immunomodulating activities [1]
The goal of this study was the generation of a recombinant monoclonal antibodies (rAbs) that binds to the beta(1,6)-branched beta-(1,3)-D-glucan schizophyllan of Schizophyllum commune
The difference to our investigations is that we proved the triple helical arrangement as an essential element for the recognition by the rAbs
Summary
A variety of fungal beta-(1,3)-D-glucans are described as substances with antitumoral and immunomodulating activities [1]. The primary molecular structure of SCH consists of a beta-(1,3)-D-glucan main chain with single beta-(1,6)-linked glucose molecules at approximately every third glucose monomer of the backbone [10, 11]. In aqueous solution it forms the secondary structure of a triplex composed of three twisted chains stabilized by hydrogen bonds with single beta-(1,6)-linked glucose residues protruding outside the helix backbone [12,13,14,15]. Aqueous solutions of SCH are viscous and show pseudoplastic flow behavior [19]. Since SCH can be used for many applications, the generation of antibodies would be of great value because they could be used for quantitative trace analysis or for the investigation of the role of particular conformations for glucan bioactivity
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