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Applied and Environmental Microbiology, December 2001, p. 5420-5424, Vol. 67, No. 12
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.12.5420-5424.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Enzyme-Linked Immunosorbent Assay Specific for (16) Branched, (13)--D-Glucan Detection in Environmental Samples

Donald K. Milton,^1,* K. Udeni Alwis,¹ Leslie Fisette,² and Michael Muilenberg¹

Department of Environmental Health, Harvard School of Public Health, Boston, Massachusetts 02115,¹ and Antibody Engineering, Eukarion, Inc., Bedford, Massachusetts 01730²

Received 4 June 2001/Accepted 14 September 2001

(13)--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 (16) branched, (13)--D-glucans in environmental samples. The assay was based on the use of a high-affinity receptor (galactosyl ceramide) specific for (13)--D-glucans as a capture reagent and a monoclonal antibody specific for fungal cell wall -D-glucans as a detector reagent. The assay was highly specific for (16) branched, (13)--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 (13)--D-glucans in house dust samples. Metal working fluids spiked with (13)--D-glucans inhibited the glucan assay. Because the assay is specific for (16) branched, (13)--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.

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^* Corresponding author. Mailing address: Occupational Health Program, Harvard School of Public Health, 665 Huntington Ave., Boston, MA 02115. Phone: (617) 432 3324. Fax: (617) 432 0219. E-mail: dmilton{at}hohp.harvard.edu.

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Applied and Environmental Microbiology, December 2001, p. 5420-5424, Vol. 67, No. 12
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.12.5420-5424.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

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