Antibody
a technology of antifungal agents and antibodies, applied in the field of antifungal antibodies, can solve the problems of not being able to detect the ia caused, compromising the ability to select the most appropriate antifungal agent,
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example 1
Fungal Culture
[0120]All fungi were cultured on Sabouraud agar (SA) under a 16 h fluorescent light regime.
Development of mAb, Preparation of Immunogen, and Immunization Regime
[0121]Mice were immunized with lyophilized mycelium (LM) of A. fumigatus AF293. Minimal medium (19 mM (NH4)2PO4, 0.5% (wt / vol) yeast extract, 7 mM sodium citrate, 2 mM MgSO4.7H2O, 0.5 mM CaCl2.2H2O and 50 mM glucose, adjusted to pH 5.5 with 1 N HCl) was sterilized by autoclaving at 121° C. for 15 min. Three-wk-old SA Petri dish cultures of the fungus were flooded with 20 ml dH2O and the conidia suspended by gentle agitation using an inoculation loop. Spore suspensions were filtered through Miracloth to remove mycelium and the filtrate containing conidia transferred to 1.5 ml micro-centrifuge tubes. The conidia were washed three times with dH2O by repeated vortexing and centrifugation at 12 000 g for 5 min and finally suspended in dH2O to give a concentration of 106 conidia / ml solution. Flasks containing 150 ml o...
example 2
Summary
[0138]Lectin binding studies show that the antigen(s) bound by MAb JF5 is / are immunogenic N-linked mannoprotein(s) comprising terminal non-reducing mannose residues linked α1-3 and α1-6. Insensitivity of the antigen(s) in ELISA to mild alkaline hydrolysis (β-elimination) shows that the MAb does not bind to glycan structures O-linked through serine and threonine.
Methodology
[0139]Lectin binding studies. Antigen(s) were purified from Aspergillus fumigatus using the method described. Purified antigen solution was subjected to glycoprotein fractionation using a Qproteome Mannose Glycoprotein Kit (Catalog no. 37551; Qiagen Ltd., Crawley, UK) according to the manufacturer's instructions. The ConA, GNA, and LCH lectin spin columns in the kit allow specific enrichment of glycoproteins with mannose-rich glycan moieties. The three lectins each bind different subclasses of these moieties. ConA binds biantennary and triantennary complex type N-glycans; LCH binds biantennary and triantenna...
example 3
Detection of Invasive Pulmonary Aspergillosis by Lateral Flow Technology Compared to Galactomannan and (1-*3)-β-D-Glucan
[0145]Early diagnosis of invasive aspergillosis is critical for the initiation of appropriate antifungal therapy and may improve outcomes in high-risk patients. The use of sensitive biomarkers, including the non-invasive assays for galactomannan and (1→3)-β-D-glucan, also reduces the use of unnecessary antifungal agents. Despite their advantages, the galactomannan and the (1→3)-β-D-glucan assays are confined to laboratories equipped for these tests or require samples be sent to reference laboratories. Lateral-flow technology incorporates immunochromatographic assays into simple devices for point-of-care diagnosis. When coupled to a monoclonal antibody specific to an extracellular glycoprotein of Aspergillus this technology is a sensitive and specific biomarker (Thornton, C. R. 2008. Development of an Immunochromatographic Lateral-Flow Device for Rapid Serodiagnosis...
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