An amplified sandwich EIA for the measurement of soy aeroallergens.
BACKGROUND
Soy hull low-molecular-weight (SHLMW) allergens were responsible for the soy asthma epidemics in Barcelona, with one 7.5 kDa protein (Gly m 1) being the main IgE-binding component. The aims of this study were to develop a sensitive sandwich enzyme immunoassay (EIA) using rabbit polyclonal antibodies to measure low levels of SHLMW allergens, and to compare this method with the previously described human IgE EIA-inhibition technique.


METHODS
IgG was isolated from serum of rabbits immunized with a chromatographically purified SHLMW extract (SHLMWE). Antibody-binding profiles were compared with those of human IgE anti-soy protein antibodies by Western blot analysis. An amplified sandwich EIA was developed using the purified SHLMWE as a calibration standard. Results were expressed in nanograms per millilitre. To compare the two assays, 54 air samples were analysed by both methods.


RESULTS
SDS-PAGE of the SHLMWE revealed four bands of 6, 8, 15 and 17 kDa. Gly m 1 in the SHLMWE was identified by fingerprinting. The detection limit of the assay was 40 pg/mL. The two methods correlated well (r=0.89; P<0.001). The allergen concentration was detected in all 54 (100%) samples by the sandwich EIA but in only 37 (68.5%) by the EIA inhibition.


CONCLUSIONS
The amplified sandwich EIA for SHLMW components has a high sensitivity and appeared to be a useful tool for the measurement of airborne SHLMW allergens, even at relatively low concentrations. Moreover, the method uses rabbit antibodies at high dilutions and does not require human sera, with limited availability and quantitative and qualitative pool-to-pool variability.
Gómez-Ollés
S
S
Pneumology and Clinic Laboratories (Immunology Unit), Hospital Vall d'Hebron, Universidad Autónoma de Barcelona, Barcelona, Spain.
Cruz
M J
MJ
Renström
A
A
Doekes
G
G
Morell
F
F
Rodrigo
M J
MJ
D000393:Air Pollutants / Q000276:immunology*; D000818:Animals; D000906:Antibodies / Q000276:immunology; D052179:Antigens, Plant / Q000276:immunology*; D019921:Biotinylation / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005512:Food Hypersensitivity / Q000276:immunology; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D007073:Immunoglobulin E / Q000276:immunology; D008970:Molecular Weight; D010936:Plant Extracts / Q000276:immunology; D011817:Rabbits; D013025:Glycine max / Q000276:immunology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D000393:Air Pollutants; D000906:Antibodies; D052179:Antigens, Plant; D010936:Plant Extracts; D007073:Immunoglobulin E