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A murine monoclonal antibody with broad specificity for occupationally relevant diisocyanates.

Authors
Lemons-AR; Siegel-PD; Mhike-M; Law-BF; Hettick-JM; Bledsoe-TA; Nayak-AP; Beezhold-DH; Green-BJ
Source
J Occup Environ Hyg 2014 Feb; 11(2):101-110
NIOSHTIC No.
20043667
Abstract
Diisocyanates (dNCOs) used in industrial applications are well known low molecular weight allergens. Occupational exposure is associated with adverse health outcomes including allergic sensitization and occupational asthma. In this study, we report the production and initial characterization of a dNCO-hapten specific murine IgM monoclonal antibody (mAb). Female BALB/c mice were immunized intraperitoneally with 25 microg of 4,4-methylene diphenyl diisocyanate (MDI)-keyhole limpet hemocyanin. Following six biweekly booster immunizations, splenocytes were recovered and fused to Sp2/0-Ag14 murine myeloma cell line for hybridoma production. Hybridomas were then screened in a solid-phase indirect enzyme-linked immunosorbent assay (ELISA) against 40:1 4,4-MDI- human serum albumin (HSA). mAb reactivity to dNCO-HSA conjugates and dNCO-HSA spiked human serum were characterized using a sandwich ELISA. One hybridoma produced a multimeric IgM mAb (15D4) that reacted with 4,4-MDI-HSA. Sandwich ELISA analysis demonstrated comparable reactivity with other occupationally relevant dNCO-HSA adducts, including 2,4-toluene diisocyanate (TDI)-HSA, 2,6-TDI-HSA, and 1,6-hexamethylene diisocyanate (HDI)-HSA, but not other electrophilic chemical HSA conjugates. The limit of quantification (LOQ) of 4,4-MDI-HSA, 2,4-TDI-HSA, 2,6-TDI-HSA, and 1,6-HDI-HSA sandwich ELISAs were 567.2, 172.7, 184.2, and 403.5ng/mL (8.67, 2.60, 2.77, and 6.07 pmol/mL), respectively. In contrast, experiments using dNCO-supplemented human sera showed an increase in the detectable limit of the assay. A mAb has been produced that has potential utility for detecting mixed diisocyanate exposures in occupational environments. The mAb may have additional utility in the standardization of specific IgE detection immunoassays as well as chromatographic-mass spectrometric methods to enrich dNCO adducted HSA in the plasma of occupationally exposed workers.
Keywords
Isocyanates; Allergens; Animal-studies; Animals; Laboratory-animals; Laboratory-testing; Exposure-assessment; Exposure-methods; Exposure-levels; Antibody-response; Immunological-tests; Enzyme-activity; Bioassays; Serological-techniques; Splenic-tissue; Author Keywords: diisocyanate; monoclonal antibody; occupational asthma; immunoassay
Contact
Brett James Green, 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
CODEN
JOEHA2
CAS No.
101-68-8; 584-84-9; 91-08-7; 822-06-0
Publication Date
20140201
Document Type
Journal Article
Email Address
dox6@cdc.gov
Fiscal Year
2014
NTIS Accession No.
NTIS Price
Identifying No.
M012014
Issue of Publication
2
ISSN
1545-9624
NIOSH Division
HELD
Source Name
Journal of Occupational and Environmental Hygiene
State
WV
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