Characterization of methylene diphenyl diisocyanate-haptenated human serum albumin and hemoglobin

Details

• Personal Author:
  Chipinda I ; Green, Brett J. ; Hettick JM ; Lemmons A ; Mhike M ; Siegel, Paul D. ; Simoyi RH
• Description:
  Protein haptenation by polyurethane industrial intermediate methylene diphenyl diisocyanate (MDI) is thought to be an important step in the development of diisocyanate (dNCO)-specific allergic sensitization; however, MDI haptenated albumins used to screen specific antibody are often poorly characterized. Recently, the need to develop standardized immunoassays using a consistent, well characterized dNCO-haptenated protein to screen for the presence of MDI-specific IgE and IgG from workers' sera has been emphasized and recognized. This has been challenging to achieve due to the bivalent, electrophilic nature of dNCO leading to the capability to produce multiple cross-linked protein species and polymeric additions to proteins. In the present study, MDI was reacted with human serum albumin (HSA) and hemoglobin (Hb) at molar ratios ranging from 1:1 to 40:1 MDI: protein. Adducts were characterized by (1) loss of available trinitrobenzene sulfonic acid (TNBS) binding to primary amines, (2) electrophoretic migration in polyacrylamide gels, (3) quantification of methylene diphenyl diamine following acid hydrolysis and (4) immunoassay. Concentration dependent changes in all the above noted parameters were observed demonstrating increase in both number and complexity of conjugates formed with increasing MDI concentration. In conclusion, a series of bio-analytical assays should be performed to standardize MDI-antigen preparations across lots and laboratories for measurement of specific antibody in exposed workers which in total indicate degree of intra- and inter-molecular cross-linking, number of dNCO bound, number of different specific binding sites on the protein and degree of immuno-reactivity. [Description provided by NIOSH]
• Subjects:
  Asthma Asthma, Occupational Chemistry Hematologic Tests Hypersensitivity Men Occupational Health Respiratory Tract Diseases Safety Women Workplace

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• Keywords:
  Allergens Allergies Author Keywords: Occupational Asthma Bronchial-asthma Chemical-properties Chemical-reactions Hemoglobin Humans Human Serum Albumin Immunoassay Standardization Methylene Diphenyl Diisocyanate Proteins Pulmonary-system-disorders Respiratory-system-disorders Work-environment Workers

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• ISSN:
  0003-2697
• 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:
  197-204
• Volume:
  440
• Issue:
  2
• NIOSHTIC Number:
  nn:20042789
• Citation:
  Anal Biochem 2013 Sep; 440(2):197-204
• Contact Point Address:
  Paul D. Siegel, Ph.D., HELD/NIOSH/CDC, 1095 Willowdale Rd., Morgantown WV 26505-2888
• Email:
  pds3@cdc.gov
• CAS Registry Number:
  Diphenylmethane diisocyanate (CAS RN 101-68-8)
• Federal Fiscal Year:
  2013
• NORA Priority Area:
  Healthcare and Social Assistance
• Peer Reviewed:
  True
• Source Full Name:
  Analytical Biochemistry
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:8bcc019bfc4813c566d6b0bdf6a8d2eacc3edea904613f7941bccb8efb5654e20b51cf7b40bd440dfc868d5d469317b9b1cd2ae96c6c458316e333ef35779826
• Download URL:
  https://stacks.cdc.gov/view/cdc/194905/cdc_194905_DS1.pdf
• File Type:
  [PDF - 711.76 KB ]