Rapid and Efficient Purification of Functional Collectin-12 and Its Opsonic Activity against Fungal Pathogens

Figure 3

Functional analysis of purified CL-12. Binding of purified CL-12 (6.5 μg/ml) to A. fumigatus was assessed by flow cytometry (a) or fluorescence microscopy (scale bar, 100 μm; 40x magnification) (b). CL-12-induced agglutination was assessed by a change of forward and side scatter morphology and shown in the inset in (a). (c) Dose-dependent binding of purified CL-12 to A. fumigatus. (d, e) Amplification of complement activation by purified CL-12. A. fumigatus were incubated with purified CL-12 (6.5 μg/ml) prior to inducing complement activation by serum^MBL- in the presence of 5 mM Mg²⁺/10 mM EGTA, and then C3b (d) or TCC (e) deposition was analyzed. The mean fluorescence intensity (MFI) was used to assess protein binding. The results are expressed as the of at least three independent experiments. .

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