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首页> 外文期刊>Annals of work exposures and health. >A Sensitive Assay for Proteases in Bioaerosol Samples: Characterization and Quantification of Airborne Proteases in Salmon Industry Work Environments
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A Sensitive Assay for Proteases in Bioaerosol Samples: Characterization and Quantification of Airborne Proteases in Salmon Industry Work Environments

机译：对生物美感样品中蛋白酶的敏感测定法：鲑鱼行业工作环境中空气中蛋白酶的表征和定量

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Proteases are probably underestimated exposure agents in bioaerosols. Their roles as barrier disrupters in allergic sensitization and activators of innate inflammation call for more attention in exposure-response studies.The main objectives of this study was (i) to establish a suitable method for detection of small quantities of proteases in filtered air samples and (ii) to utilize the method to characterize exposure to proteases in a salmon industry work environment. Analysis of proteases in filtered air samples was based on zymography, using sodium dodecyl sulfate-polyacrylamide gel electrophoresis with 0.1% gelatin as substrate added in the polyacrylamide gel. Gelatinase activity was evident as cleared (unstained) regions.The area of these regions was quantified using image analysis (UVP Vision Works (R)). Standard curves with known amounts of active porcine trypsin were added to each gel. Validation of 11 non-linear standard curves showed R-2 (range) = 0.8989-0.9882, limit of detection = 0.056 nM, lower limit of quantification = 0.161 nM, and coefficients of variations (range) = 20-28%. Sampling of bioaerosols in salmon industry was performed using polytetrafluoretylene filters with an airflow of 31 min(-1). All samples contained visible bands close to the size of porcine trypsin (23.3 kDa). The bands did not disappear in the presence of EDTA but abolished by Pefabloc, demonstrating that the enzyme is a serine protease, most likely salmon trypsin. Airborne levels of active protease were below the statistical detection limit in the filleting department but quantifiable in extract samples from the slaughter department. Three filtered air samples from the slaughter department showed air concentrations of 6.2, 16.5, and 27.0 ng m(3) air. We conclude that zymography is a sensitive and reliable method for exposure assessment of active proteases in indoor environmental samples. We recommend this assay for use in occupational studies to characterize and quantify exposure to active proteases in bioaerosols.
机译：蛋白酶可能低估了生物溶质中的暴露剂。它们在过敏反应中作为障碍的作用和先天炎症激活剂在暴露响应研究中需要更多注意。这项研究的主要目标是（i）建立一种适合检测过滤空气样品和少量蛋白酶的方法（ii）利用该方法来表征鲑鱼行业工作环境中蛋白酶的暴露。使用十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳，用0.1％明胶添加在聚丙烯酰胺凝胶中的底物，对过滤的空气样品中的蛋白酶进行了基于Zymography的分析。明显明显的是明显的（未染色）区域。使用图像分析（UVP Vision Works（R））对这些区域的面积进行了量化。将具有已知量的活性猪胰蛋白酶的标准曲线添加到每个凝胶中。 11个非线性标准曲线的验证显示R-2（范围）= 0.8989-0.9882，检测极限= 0.056 nm，量化下限= 0.161 nm，变化系数（范围）= 20-28％。使用Polytetrafletrafluoretylene过滤器进行的气流（-1）进行了鲑鱼行业中生物溶质的采样。所有样品均包含接近猪胰蛋白酶（23.3 kDa）的可见条带。在EDTA存在的情况下，乐队没有消失，而是被Pefabloc废除，表明该酶是一种丝氨酸蛋白酶，很可能是鲑鱼胰蛋白酶。空气中的活性蛋白酶水平低于圆角部门的统计检测极限，但可以在屠宰部的提取样品中进行量化。屠宰部门的三个过滤空气样品显示空气浓度为6.2、16.5和27.0 ng m（3）空气。我们得出的结论是，酶机是一种敏感且可靠的方法，用于对室内环境样品中活性蛋白酶的暴露评估。我们建议该测定法用于职业研究，以表征和量化生物溶质中活性蛋白酶的暴露。

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