Enhancement of murine IgE antibody detection by IgG removal.
BACKGROUND
Although animal models for the study of allergic reactions are desirable, the use of mice has been hindered by the lack of sufficiently sensitive in vitro immunoglobulin epsilon (IgE) antibody assays. The aim of this study was to enhance IgE antibody measurements by immunoglobulin gamma (IgG) depletion.


METHODS
Seven- to eight-week-old female mice of four strains (C3H/HeJ, CBA/J, C57Bl/6J, and Balb/c) were immunized (20 mice/group) with shrimp or peanut extracts using Al(OH)(3) as adjuvant. Following immunization, animals were sacrificed by exsanguination and the sera of each group pooled. Initial measurements of IgE antibody levels by enzyme-linked immunosorbent assay (ELISA) were relatively low; IgG and IgE reactivity patterns by immunoblot were similar. Thus, sera from shrimp or peanut immunized mice were depleted of IgG (absorbed 3-6 times with immobilized protein G) and then tested for IgE antibody to shrimp or peanut allergen.


RESULTS
A 3- to 5-fold increase in IgE antibody reactivity as measured by ELISA was demonstrated when >80-90% of the IgG was removed. This increase in detection of allergen-specific IgE occurred in sera from all mouse strains and to all allergens tested. In addition, reactivity of IgE antibodies to peanut or shrimp allergens by immunoblot increased visually approximately 4- to 10-fold.


CONCLUSIONS
These studies indicate that allergen-specific IgG antibodies, which may be in more than 100-fold excess to IgE antibodies, interferes with detection of allergen-specific IgE, probably by competitive binding to allergenic epitopes. Substantial depletion of IgG antibodies (>80%) result in a significant increase in the sensitivity of the antibody measurements.
Lehrer
S B
SB
Tulane University Health Sciences Center, 1700 Perdido Street (SL-57), New Orleans, LA 70112, USA. sblehrer@tulane.edu
Reish
R
R
Fernandes
J
J
Gaudry
P
P
Dai
G
G
Reese
G
G
D000485:Allergens; D000818:Animals; D052179:Antigens, Plant; D010367:Arachis / Q000276:immunology; D060829:Arthropod Proteins; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006023:Glycoproteins; D007073:Immunoglobulin E / Q000097:blood* / Q000276:immunology; D007074:Immunoglobulin G / Q000097:blood* / Q000276:immunology; D008565:Membrane Proteins; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D009419:Nerve Tissue Proteins / Q000502:physiology; D010940:Plant Proteins / Q000276:immunology; D011506:Proteins / Q000276:immunology
D000485:Allergens; D052179:Antigens, Plant; C500889:Ara h 1 protein, Arachis hypogaea; D060829:Arthropod Proteins; C029152:G-substrate; D006023:Glycoproteins; D007074:Immunoglobulin G; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D010940:Plant Proteins; D011506:Proteins; C058449:shrimp allergens Sa; D007073:Immunoglobulin E