PMC7734872_01
Proteins extracted from four metalworking fluid (MWF) samples. A: Silver stain of SDS-PAGE; B: sIgG blot with serum of MWF-exposed patient; C: sIgG blot with non-exposed reference serum. Lane 1 - 4 proteins from MWF pellet fraction (MWF-13 - MWF-16), lane 6 - 8 proteins from MWF supernatant fraction (MWF-13 - MWF-16).
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Figure1
All four MWF workplace samples (MWF pellet fraction and MWF supernatant fraction) contained antigens that were visible in silver-stained gel electrophoresis in the range of 5 - 100 kDa (Figure 1A).  C). Clear IgG binding to antigens was seen with serum of the MWF-exposed patient (Figure 1B), with dominant markers of the protein bands at 10, 15, 27, 60, and 100 kDa and further protein bands in the pellet and supernatant fractions. The reference serum tested for the same protein extracts (Figure 1C) showed only weak IgG bands on proteins of the MWF pellet fraction.
Proteins extracted from metalworking fluid (MWF) bacterial isolates. A: Silver stain of SDS-PAGE; B: sIgG blot with serum of MWF exposed patient; C: sIgG blot with non-exposed reference serum. lane 1: Pseudomonas oleovorans, lane 2: Pseudomonas alcaliphila, lane 3: Pseudomonas spec., lane 4: Paenibacillus glucanolyticus, lane 5: Corynebacterum amycolatum.
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Figure2
The protein spectra of the bacteria were individually different in the molecular weight range of 5 - 100 kDa (Figure 2A). A qualitative IgG detection of the bacterial antigens by immunoblot with the serum of the MWF-exposed patient showed a significantly stronger sIgG antigen binding compared to a reference serum (Figure 2 B, C).