A Sensitive Assay for Proteases in Bioaerosol Samples: Characterization and Quantification of Airborne Proteases in Salmon Industry Work Environments
| Autor: | Sampada Satchidanand Bhagwat, Berit Bang, Nabin Malla, Jan-Olof Winberg, Lisbeth Aasmoe |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Proteases medicine.medical_treatment Indoor bioaerosol Fisheries Air Pollutants Occupational 03 medical and health sciences 0302 clinical medicine Salmon Occupational Exposure Endopeptidases medicine Animals Humans Zymography Serine protease Detection limit Aerosols Protease Chromatography biology Chemistry Public Health Environmental and Occupational Health Trypsin 030210 environmental & occupational health 030104 developmental biology biology.protein Biological Assay Electrophoresis Polyacrylamide Gel medicine.drug Bioaerosol Peptide Hydrolases |
| Zdroj: | Annals of work exposures and health. 62(8) |
| ISSN: | 2398-7316 |
| Popis: | 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®). Standard curves with known amounts of active porcine trypsin were added to each gel. Validation of 11 non-linear standard curves showed R2 (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 3 l 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|