Abstract
There is a real medical need of new diagnostic tools for the early recognition of invasive Candida infections. We exploited a rather simple and rapid redox methodology to construct a bispecific monoclonal antibody (bsmAb) that combines a monoclonal antibody (mAb) directed against 1,3-β-D-glucan, a well-known, pan-fungal diagnostic biomarker, with a mAb recognizing MP65, a major immunogenic mannoprotein secreted by C.albicans and other Candida species. The bsmAb (MP65/bglu mAb) was successfully produced and purified at high yields and proved to bind and reveal simultaneously, with high sensitivity, the β-glucan and MP65 antigens in both purified and native forms. The MP65/bglu mAb is the first bispecific antibody generated against a fungal microorganism and may prove useful for the concurrent detection of different and clinically significant Candida biomarkers in patient sera.
Highlights
Invasive fungal infections caused by C. albicans and other Candida species are a major cause of morbidity and mortality in hospitalized patients worldwide [1,2]
The second monoclonal antibody (mAb) (4C8) is a mouse IgG1 that recognizes the protein moiety of MP65 (MP65p), a highly immunogenic mannoprotein involved in Candida cell wall maintenance and remodeling [15,16,24]
BsmAbs proposed for the diagnosis of human microbial infections are even fewer, and were all constructed to bind in combination a microbial antigen and various types of reporter molecule, in order to improve antigen recognition specificity in different immunoassays [33,34,35]
Summary
Invasive fungal infections caused by C. albicans and other Candida species are a major cause of morbidity and mortality in hospitalized patients worldwide [1,2]. Despite the recent development and clinical availability of antifungal drugs of high efficacy and reduced toxicity, these infections remain difficult to control and their associated mortality rates are still unacceptably high, approaching 40–50% in some cases. This is further burdened by the high cost of treatment [1,2]. A number of noncultural assays, mostly based on the detection of fungal biomarkers in patient circulation, are PLOS ONE | DOI:10.1371/journal.pone.0148714. A number of noncultural assays, mostly based on the detection of fungal biomarkers in patient circulation, are PLOS ONE | DOI:10.1371/journal.pone.0148714 February 9, 2016
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