Enzyme-Linked Immunosorbent Assay Specific for (1→6) Branched, (1→3)-β- d -Glucan Detection in Environmental Samples

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Abstract
(1→3)-β- d -Glucans have been recognized as a potential causative agent responsible for bioaerosol-induced respiratory symptoms observed in both indoor and occupational environments. A specific enzyme immunoassay was developed to quantify (1→6) branched, (1→3)-β- d -glucans in environmental samples. The assay was based on the use of a high-affinity receptor (galactosyl ceramide) specific for (1→3)-β- d -glucans as a capture reagent and a monoclonal antibody specific for fungal cell wall β- d -glucans as a detector reagent. The assay was highly specific for (1→6) branched, (1→3)-β- d -glucans (such as that from Saccharomyces cerevisiae ) and did not show any response at 200 ng/ml to curdlan, laminarin, pustulan, dextran, mannan, carboxymethyl cellulose, and endotoxins. The detection level was 0.8 ng/ml for baker's yeast glucan and Betafectin. A coefficient of variation of 7.8% was obtained for (1→3)-β- d -glucans in house dust samples. Metal working fluids spiked with (1→3)-β- d -glucans inhibited the glucan assay. Because the assay is specific for (1→6) branched, (1→3)-β- d -glucans and is sensitive and reproducible, it will be useful for the investigation of health effects from exposure to this class of biologically active molecules.