Development of sandwich ELISAs for the detection of aromatic diisocyanate adducts

Details

• Personal Author:
  Beezhold, Donald H. ; Bledsoe TA ; Green, Brett J. ; Lemons, Angela R. ; Siegel, Paul D.
• Description:
  Diisocyanates (dNCOs) are highly reactive low molecular weight chemicals commonly used in the manufacturing industry. Occupational exposures to dNCOs have been shown to elicit allergic sensitization and occupational asthma. Among the most commonly used dNCOs in industry are the aromatic dNCOs, toluene diisocyanate (TDI) and methylene diphenyl diisocyanate (MDI). This study aimed to develop enzyme linked immunosorbent assays (ELISA) utilizing aromatic dNCO-specific monoclonal antibodies (mAbs) for the detection of aromatic dNCO adducts. Two sandwich ELISAs were developed. The first sandwich ELISA utilized mAb 60G2 along with an anti-human serum albumin (HSA) polyclonal antibody. This assay detected MDI-, 2,4- and 2,6-TDI-HSA adducts with limits of detection (LOD) of 2.67, <0.10, and 1.70ng/mL, respectively. When spiked into human serum, the LOD of this ELISA increased to 34.37, 7.64 and 24.06ng/mL, respectively. The second ELISA utilized mAbs 62G5 and 60G2 for capture and detection. This assay was capable of detecting 2,4- and 2,6-TDI-HSA adducts with LODs of <4.90 and 26.92ng/mL, respectively, and when spiked in human serum, <4.90 and 95.93ng/mL, respectively. This 62G5-60G2 sandwich assay was also able to detect dNCO adducted transferrin, hemoglobin, keratin and actin, but with less sensitivity than dNCO-HSA. The results of this study demonstrate potential application of these ELISAs in the identification and characterization of aromatic dNCO adducts as well as in biomonitoring occupational and environmental dNCO exposures. [Description provided by NIOSH]
• Subjects:
  Antibodies Asthma Asthma, Occupational Biochemistry Biological Assay Chemistry Enzymes Hypersensitivity Isocyanates Occupational Health Polycyclic Aromatic Hydrocarbons Safety Toluene

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• Keywords:
  Allergies Analytical-instruments Analytical-methods Antibody-response Aromatic-hydrocarbons Author Keywords: Diisocyanate Bioassays Biochemical-tests Biological-monitoring Blood-serum Bronchial-asthma Chemical-indicators Immunoassay Immunological-tests Methyl-compounds Monoclonal Antibody Occupational Asthma Phenyls Toluenes

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• ISSN:
  0022-1759
• Document Type:
  Text
• Genre:
  Journal Article
• Place as Subject:
  OSHA Region 3 ; West Virginia
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Division:
  HELD - Health Effects Laboratory Division
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Pages in Document:
  66-70
• Volume:
  397
• Issue:
  1
• NIOSHTIC Number:
  nn:20043355
• Citation:
  J Immunol Methods 2013 Nov; 397(1-2):66-70
• Contact Point Address:
  Angela R. Lemons, Allergy and Clinical Immunology Branch, Health Effects Laboratory Branch, National Institute for Occupational Safety and Health, Centers for Disease Control and Prevention, 1095 Willowdale Road, Morgantown, WV 26505, USA
• Email:
  wrw0@cdc.gov
• CAS Registry Number:
  Diphenylmethane diisocyanate (CAS RN 101-68-8) ; Toluene 2,4-diisocyanate (CAS RN 584-84-9) ; Toluene 2,6-diisocyanate (CAS RN 91-08-7) ; Toluene diisocyanate (CAS RN 26471-62-5)
• Federal Fiscal Year:
  2014
• Peer Reviewed:
  True
• Source Full Name:
  Journal of Immunological Methods
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:6f43ef968ca591def395d0e36393210e28b9aaf9f07ca6ae731970e140cf0e836ae1a3172c3f969e091d08cfecbee8e5061b21223b5038cf2b9bd79cbb817895
• Download URL:
  https://stacks.cdc.gov/view/cdc/195273/cdc_195273_DS1.pdf
• File Type:
  [PDF - 458.61 KB ]