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Specific monoclonal antibodies and human immunoglobulin E show that Hev b 5 is an abundant allergen in high protein powdered latex gloves
Author(s) -
Sutherland M. F.,
Drew A.,
Rolland J. M.,
Slater J. E.,
Suphioglu C.,
O'Hehir R. E.
Publication year - 2002
Publication title -
clinical and experimental allergy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.462
H-Index - 154
eISSN - 1365-2222
pISSN - 0954-7894
DOI - 10.1046/j.0954-7894.2002.01355.x
Subject(s) - latex allergy , maltose binding protein , polyclonal antibodies , allergen , immunoglobulin e , fusion protein , monoclonal antibody , recombinant dna , antibody , virology , allergy , microbiology and biotechnology , immunology , chemistry , biology , biochemistry , gene
Summary Background Hev b 5 is a major latex allergen recognized predominantly by latex‐allergic health care workers (HCWs). Recombinant Hev b 5 (rHev b 5) was previously expressed as a fusion protein with maltose binding protein (MBP), itself an immunogenic molecule; therefore non‐fusion rHev b 5 is desirable. Moreover, standardized immunological assays for the detection of Hev b 5 are currently lacking and may have important implications for both allergen avoidance and diagnosis in latex allergy. Objectives To generate and use Hev b 5‐specific mAbs to determine the relative abundance of Hev b 5 in different latex extracts, correlating this with the IgE reactivity of latex‐allergic HCWs and to produce non‐fusion rHev b 5. Methods For the production of mAbs, mice were immunized with rHev b 5/MBP fusion protein and mAbs selected with rHev b 5/MBP but not MBP reactivity. The mAb reactivity was compared with polyclonal IgE from latex‐allergic HCWs using direct and inhibition ELISA and immunoblot assays. Recombinant Hev b 5 was expressed and purified in the pPROEX‐HTa bacterial expression system. Results Four Hev b 5‐specific mAbs were produced. Immunoblotting and ELISA using the mAbs indicate abundant Hev b 5 in high protein powdered latex glove extracts as compared with crude latex sap extracts. High quality surgical gloves with no detectable protein have no detectable Hev b 5. Inhibition ELISAs using serum IgE from latex‐allergic HCWs and Hev b 5‐specific mAbs gave strong correlation. Non‐fusion recombinant Hev b 5 was successfully expressed and purified, showing reactivity with both the Hev b 5‐specific mAbs and serum IgE of latex‐allergic HCWs. Conclusion Hev b 5‐specific mAbs and human IgE from latex‐allergic HCWs demonstrate the greater content of Hev b 5 in high protein powdered glove extracts. This may explain the observed higher frequency of sensitization to this allergen in HCWs.