Domain mapping and comparative binding features of eight dog IgE-specific reagents in ELISA, immunoblots, and immunohistochemistry.
Eight dog IgE-specific reagents including monoclonal and polyclonal antibodies (Ab) and a cross-reactive alpha chain of the human high affinity IgE receptor were mapped to recombinant fragments of the second (IgEf2) and third/fourth (IgEf3/4) domains of the dog IgE heavy chain. In ELISA, five out of eight reagents reacted to solid-phase bound IgEf2, of which two polyclonal Ab bound in addition to IgEf3/4. All Ab which recognized at least one recombinant IgE fragment, also bound to IgE in ELISA, immunoblots, and immunohistochemistry. In contrast, only one monoclonal Ab, that did not bind to the recombinant IgE fragments, reacted with immunoblots of serum and immunohistochemistry. The alpha chain could only be applied to ELISA with serum IgE. Furthermore, there was a wide range of heat-lability of binding reactions. Comparative analysis of available dog IgE-specific reagents enables more in-depth functional studies on IgE-mediated phenomena in dogs, and helps to further establish the dog as an animal model for allergy research.
Griot-Wenk
M E
ME
Division of Immunogenetics, Institute of Animal Breeding, Switzerland. monika.griot-wenk@ch.novartis.com
Marti
E
E
DeBoer
D J
DJ
de Weck
A L
AL
Lazary
S
S
D000818:Animals; D004285:Dogs / Q000097:blood / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D018604:Epitope Mapping / Q000379:methods / Q000662:veterinary*; D006801:Humans; D007073:Immunoglobulin E / Q000032:analysis*; D007150:Immunohistochemistry; D007163:Immunosorbent Techniques / Q000662:veterinary; D007202:Indicators and Reagents; D011485:Protein Binding; D011994:Recombinant Proteins / Q000276:immunology
D007202:Indicators and Reagents; D011994:Recombinant Proteins; D007073:Immunoglobulin E