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569 Reduction in airborne latex protein exposure by use of low protein, powder-free gloves
J ALLERGYCLIN IMMUNOL VOLUME97, NUMBER1, PART3
569
Reduction i n A i r b o r n e L a t e x P r o t e i n Exposure By Use o f Low P r o t e i n , Powder-Free G l o v e s . SR Siu
Abstracts
571
In vivo and in vitro E v a l u a t i o n o f A l l e r g e n i c i t y of N a t u r a l R u b b e r L a t e x ( N R L ) Gloves u s e d in H e a l t h Care: A N a t i o n - w i d e S t u d y . K Turianmaa MD. S MltkinenKiljunen M~, H Alenius Msc. T Reunal~ MD, T Palosuo MD. Tampere and Helsinki, Finland. NRL allergy is a well-recognized problem among health care workers using protective gloves. Data has accumulated to indicate that there may be considerable differences between the allergenicity of gloves of different manofacturers. The aim of the present study was to measure NRL allergen content in a sample covering >90% of glove brands marketed in Finland. 20 brands of surgical and examination gloves were collected. Coded extracts (1:5, w/v) were blindly skin prick tested in latex-allergic patients (n=20) and assayed for total NRL allergen activity by RAST-inhibition 0LI) and a novel ELISA-inhibitlnn (EI) tesL The results between RI and El correlated highly significantly with each other (r=0.96) and with prick tests (r=-0.94-0.96) indicating that these 2 in vitro methods can be used for reliable NRL allergen measurement, thus decreasing the need for in vivo testing. Over 1000-fold differences in allergenicity of various gloves were seen. The allergen content (100 000 arbitrary tLrtits(AU)/ml in NRL) was considered high (> 100 AUhnl) in 7, moderate (100-10 AUIml) in 2, and low (<10 AU/ml) in 9 glove brands. Glove marketing companies and glove purckasing personnel in hospitals were informed of the observed wide variation in allergenieity of the gloves. It is likely that this information will have a clear impact on future glove purchasing policies in Finland.
Characterization of Latex Allergens by Capillary E l e c t r o p h o r e s i s . HI) Chen MS. CL Chen PhD. SW Huana MD. Gainesville, Florida Latex allergens were studied by capillary electrophoresis to determine whether they are composed of similar protein subnnits. The separation mechanism of capillary electrophoresis is based on differences in the charge-to-mass ratio. Capillary zone electrophoresis is the most widely used and simplest mode of capillary electrophoresis. Latex allergen components, approximately 12, 22, 30, 34, 46, and 58kD in molecular weight, were cut from SDS-PAGE gels and eluted. Eluates were concentrated with an ultrafiltration apparatus, run through Bio-R,'ld SM-2 gel to remove SDS and applied to a capillary electrophoresis apparatus with a diode array detector. Capillary zone electrophoresis separations were performed in 100 mM borate buffer, pH 9.2. Electrophoregrams showed latex allergen eompunents cut from the gels had similar migration patterns, and each gave peaks at approximately 30 and 40 minutes migration time. Diode array spectral analysis of these peaks indicated the latex components were proteins that share similar spectral characteristics. Additionally, these proteins are immunogenic; immunoglobulin from latex-allergic patients was incubated with the 12, 22, or 34kD allergen components and run under capillary zone electrophoresis conditions. An additional peak, presumedly the antibody-antigen complex, appeared on the electrophoregrams, suggesting immunoreactivity in these three latex allergen components. These results suggest that the 12, 22, 30, 34, 46, and 58 kD components of latex protein could be similar in structure, based on the criterion of charge-to-mass ratio. The proteins migrating at approximately 30 and 40 minutes by capillary zone electruphoresis will be isolated and purified for further study.
MO, GO Smith, GL Sussman MD. MC Swanson PhD, d Yunqinqer MD, MP Cividino MD, SA Brown RN, D Beezhold PhD, London, Ontario, Canada Allergic reactions to latex gloves have been attributed to exposure to airborne latex protein. This study compares airborne exposure levels during the use of regular latex gloves and low protein, powder-free latex gloves. Both personal and area samples were taken OR the wards and in the operating rooms of two hospitals during summer and winter. Latex allergens were quantified by an inhibition immunoassay using latex-specific human IgE. For personal sampling, the results of control gloves (12 samples) ranged between 5 and 616 ng/M3 with a mean of 119 ng/M . When low protein, powder-free gloves were used, none of the 14 personal samples showed any detectable airborne protein. Area samples showed similar results but were of lower concentration (4 to 33 ng/M3 for control gloves and less than 0.5 ng/M° for low protein, powder-free gloves). Exposure levels were higher during the winter, likely due to reduced fresh air input. This study demonstrates that airborne exposure to latex protein can be reduced by the use of low protein powder-free gloves instead of regular latex gloves.
570
325
572
Electrophoretic characterization of latex a l l e r g e n s b y two-dimensional e l e c t r o p h o r e s i s . A P o s c h PhD. Z Chen PhD, M Raulf-Heirosoth PhD. X Baur MD. B o c h u m , Germany The latex sap of Hevea brasiliensis contains a complex protein mixture responsible for IgE-mediated latex hypersensitivity. We have applied high resolution two-dimensional (2-D) electrophoresis with immobilized pH gradients (IPG-Dalt) to separate latex proteins and to subsequently identify those components that bind IgE by immunoblotting. Both the analytical (80 lag protein load) and the micropreparative (400 lag) IPG-Dalt analyses (1. dimension: IPG 4-8; 2. dimension: SDS-PAGE, T= 12%) were highly reproducible and showed excellently resolved spots without streaking. More than 350 single protein spots could be detected on the silver stained analytical gels. The most distinct 150 spots were also visible on the Coomassie-stained micropreparative 2-D blots using polyvinylidene difluoride (PVDF) membranes. Major IgE-binding was detected in the 46, 30, 20, l 5 and <10 kD area of the micropreparative 2-D immunoblots. Our results indicate that IPG-DALT is due to its improved reproducibility and high protein loading capacity the method of choice for the micropreparative isolation of IgE binding latex proteins. Moreover, in combination with blotting onto an inert support matrix which can be directly applied to an automated protein sequencer or Matrix Assisted Laser Desorption-MS, we are now able to characterize simultaneously a variety of different latex allergens in a very short period of time.
569
Reduction i n A i r b o r n e L a t e x P r o t e i n Exposure By Use o f Low P r o t e i n , Powder-Free G l o v e s . SR Siu
Abstracts
571
In vivo and in vitro E v a l u a t i o n o f A l l e r g e n i c i t y of N a t u r a l R u b b e r L a t e x ( N R L ) Gloves u s e d in H e a l t h Care: A N a t i o n - w i d e S t u d y . K Turianmaa MD. S MltkinenKiljunen M~, H Alenius Msc. T Reunal~ MD, T Palosuo MD. Tampere and Helsinki, Finland. NRL allergy is a well-recognized problem among health care workers using protective gloves. Data has accumulated to indicate that there may be considerable differences between the allergenicity of gloves of different manofacturers. The aim of the present study was to measure NRL allergen content in a sample covering >90% of glove brands marketed in Finland. 20 brands of surgical and examination gloves were collected. Coded extracts (1:5, w/v) were blindly skin prick tested in latex-allergic patients (n=20) and assayed for total NRL allergen activity by RAST-inhibition 0LI) and a novel ELISA-inhibitlnn (EI) tesL The results between RI and El correlated highly significantly with each other (r=0.96) and with prick tests (r=-0.94-0.96) indicating that these 2 in vitro methods can be used for reliable NRL allergen measurement, thus decreasing the need for in vivo testing. Over 1000-fold differences in allergenicity of various gloves were seen. The allergen content (100 000 arbitrary tLrtits(AU)/ml in NRL) was considered high (> 100 AUhnl) in 7, moderate (100-10 AUIml) in 2, and low (<10 AU/ml) in 9 glove brands. Glove marketing companies and glove purckasing personnel in hospitals were informed of the observed wide variation in allergenieity of the gloves. It is likely that this information will have a clear impact on future glove purchasing policies in Finland.
Characterization of Latex Allergens by Capillary E l e c t r o p h o r e s i s . HI) Chen MS. CL Chen PhD. SW Huana MD. Gainesville, Florida Latex allergens were studied by capillary electrophoresis to determine whether they are composed of similar protein subnnits. The separation mechanism of capillary electrophoresis is based on differences in the charge-to-mass ratio. Capillary zone electrophoresis is the most widely used and simplest mode of capillary electrophoresis. Latex allergen components, approximately 12, 22, 30, 34, 46, and 58kD in molecular weight, were cut from SDS-PAGE gels and eluted. Eluates were concentrated with an ultrafiltration apparatus, run through Bio-R,'ld SM-2 gel to remove SDS and applied to a capillary electrophoresis apparatus with a diode array detector. Capillary zone electrophoresis separations were performed in 100 mM borate buffer, pH 9.2. Electrophoregrams showed latex allergen eompunents cut from the gels had similar migration patterns, and each gave peaks at approximately 30 and 40 minutes migration time. Diode array spectral analysis of these peaks indicated the latex components were proteins that share similar spectral characteristics. Additionally, these proteins are immunogenic; immunoglobulin from latex-allergic patients was incubated with the 12, 22, or 34kD allergen components and run under capillary zone electrophoresis conditions. An additional peak, presumedly the antibody-antigen complex, appeared on the electrophoregrams, suggesting immunoreactivity in these three latex allergen components. These results suggest that the 12, 22, 30, 34, 46, and 58 kD components of latex protein could be similar in structure, based on the criterion of charge-to-mass ratio. The proteins migrating at approximately 30 and 40 minutes by capillary zone electruphoresis will be isolated and purified for further study.
MO, GO Smith, GL Sussman MD. MC Swanson PhD, d Yunqinqer MD, MP Cividino MD, SA Brown RN, D Beezhold PhD, London, Ontario, Canada Allergic reactions to latex gloves have been attributed to exposure to airborne latex protein. This study compares airborne exposure levels during the use of regular latex gloves and low protein, powder-free latex gloves. Both personal and area samples were taken OR the wards and in the operating rooms of two hospitals during summer and winter. Latex allergens were quantified by an inhibition immunoassay using latex-specific human IgE. For personal sampling, the results of control gloves (12 samples) ranged between 5 and 616 ng/M3 with a mean of 119 ng/M . When low protein, powder-free gloves were used, none of the 14 personal samples showed any detectable airborne protein. Area samples showed similar results but were of lower concentration (4 to 33 ng/M3 for control gloves and less than 0.5 ng/M° for low protein, powder-free gloves). Exposure levels were higher during the winter, likely due to reduced fresh air input. This study demonstrates that airborne exposure to latex protein can be reduced by the use of low protein powder-free gloves instead of regular latex gloves.
570
325
572
Electrophoretic characterization of latex a l l e r g e n s b y two-dimensional e l e c t r o p h o r e s i s . A P o s c h PhD. Z Chen PhD, M Raulf-Heirosoth PhD. X Baur MD. B o c h u m , Germany The latex sap of Hevea brasiliensis contains a complex protein mixture responsible for IgE-mediated latex hypersensitivity. We have applied high resolution two-dimensional (2-D) electrophoresis with immobilized pH gradients (IPG-Dalt) to separate latex proteins and to subsequently identify those components that bind IgE by immunoblotting. Both the analytical (80 lag protein load) and the micropreparative (400 lag) IPG-Dalt analyses (1. dimension: IPG 4-8; 2. dimension: SDS-PAGE, T= 12%) were highly reproducible and showed excellently resolved spots without streaking. More than 350 single protein spots could be detected on the silver stained analytical gels. The most distinct 150 spots were also visible on the Coomassie-stained micropreparative 2-D blots using polyvinylidene difluoride (PVDF) membranes. Major IgE-binding was detected in the 46, 30, 20, l 5 and <10 kD area of the micropreparative 2-D immunoblots. Our results indicate that IPG-DALT is due to its improved reproducibility and high protein loading capacity the method of choice for the micropreparative isolation of IgE binding latex proteins. Moreover, in combination with blotting onto an inert support matrix which can be directly applied to an automated protein sequencer or Matrix Assisted Laser Desorption-MS, we are now able to characterize simultaneously a variety of different latex allergens in a very short period of time.