An ELISA-based method for measurement of food-specific IgE antibody in mouse serum: an alternative to the passive cutaneous anaphylaxis assay.
Passive cutaneous anaphylaxis (PCA) assay has been a gold standard method to measure allergen-specific IgE antibody (ASIgE Ab) levels in allergy mouse models. Many factors including stringent guidelines for laboratory animal use make PCA a difficult choice. Therefore, alternative methods are needed that can be readily applied for measurement of specific IgE antibody levels in mouse serum. Herein we describe a novel ELISA-based method that is more sensitive in comparison to PCA, IgE isotype-specific (because it has little cross-reactivity with IgG1 or IgG2a isotype) and highly reproducible (<10% inter- or intra-assay variation). Furthermore, we demonstrate the utility of this assay to measure specific IgE Ab against a variety of food extracts including chicken egg, peanut, almond, filbert/hazelnut and sweet potato. These findings are of particular interest to those who are seeking (i) to measure food-extract-specific IgE antibody in animal models and (ii) an alternative to the animal-based PCA method to measure mouse IgE antibodies.
Birmingham
Neil
N
Food Allergy and Immunology Laboratory, Department of Food Science and Human Nutrition, Michigan State University, East Lansing, MI 48823, USA.
Payankaulam
Sandhya
S
Thanesvorakul
Sirinart
S
Stefura
Bill
B
HayGlass
Kent
K
Gangur
Venu
V
D000485:Allergens; D000818:Animals; D000918:Antibody Specificity; D003429:Cross Reactions; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000706:statistics & numerical data; D005502:Food / Q000009:adverse effects; D005512:Food Hypersensitivity / Q000276:immunology*; D007073:Immunoglobulin E / Q000032:analysis* / Q000097:blood; D007074:Immunoglobulin G / Q000032:analysis / Q000097:blood; D007139:Immunoglobulin epsilon-Chains / Q000032:analysis / Q000097:blood; D051379:Mice; D010323:Passive Cutaneous Anaphylaxis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000485:Allergens; D007074:Immunoglobulin G; D007139:Immunoglobulin epsilon-Chains; D007073:Immunoglobulin E