Pediatr Allergy Respir Dis.  2000 Mar;10(1):34-40.

Immunoblot Analysis of Hypoallergenic Buckwheat with Monoclonal Antibodies to Raw Buckwheat

• Yum HY
• Ryu JW
• Kim KE
• Lee KY

Abstract

BACKGROUND AND OBJECTIVE
It is generally accepted that one third of children with food allergy lose their clinical activity if the responsible food allergen can be identified and completely eliminated from their diet. But buckwheat allergy rarely lose clinical activity. So, we have produced hypoallergenic buckwheat(HBW) according to Ikezawa's method, and characterized the allergenicity by immunoblot method using monoclonal antibodies to raw buckwheat. MATERIAL AND METHODS: HBW was prepared from raw buckwheat by salting with 0.5 M NaCl. 18 monoclonal antibodies to raw buckwheat were made according to standard polyehtylene glycol method. We performed SDS-PAGE of HBW and transferrd to nitrocellulose membranes, which were immunoblotted by selected 3 monoclonal antibodies to raw buckwheat.
RESULTS
The protein concentration of HBW was 47.3 microgram/microliter. As the result of SDS-PAGE of HBW the protein bands of 50, 36, 13 kD of raw buckwheat disappeared. 3 monoclonal antibodies were selected due to their higher specificities in ELISA. Immunoblot study showed that 2 among 3 monoclonal antibodies bind to HBW.
CONCLUSION
The SDS-PAGE of HBW showed less protein bands in comparison to raw buckwheat. But the allergic component of HBW still remained. So further studies are required to eliminate the residual allergenicity of hypoallergenic food.

Keyword

Hypoallergenic buckwheat; Immunoblot; Monoclonal antibody

MeSH Terms

Antibodies, Monoclonal*
Child
Collodion
Diet
Electrophoresis, Polyacrylamide Gel
Enzyme-Linked Immunosorbent Assay
Fagopyrum*
Food Hypersensitivity
Humans
Hypersensitivity
Membranes
Antibodies, Monoclonal
Collodion