Covalent-binding native blot in the determination of specific IgE to latex gloves allergens. Comparison with other methods.
Six sera from patients with clinical symptoms of latex hypersensitivity and latex-RAST values ranging from class 1 to 4 were used to characterize latex-glove allergens by immunoblotting. Twelve % acrylamide-SDS-PAGE of the latex glove crude extract revealed a single band with an estimated mol wt lower than 14,000 D. The proteins bound to nitrocellulose after electrical transfer as shown by Ponceau red staining of the membrane. However, standard immunoblotting methods using alkaline-phosphatase anti-IgE did not detect the presence of specific IgE. Different variants of SDS-PAGE immunoblotting were also unsuccessful. On the other hand, the use of IEF and capillary transfer (native blot) to a covalent-binding hydrophilic membrane was found to be the method of choice for the study of this allergen. The authors believe that this procedure can be helpful in the study of other antigen-antibody systems, particularly when standard immunoblotting assays fail.
Moneo
I
I
Servicio de Biopatología, Instituto de Salud Carlos III, Madrid.
Nocito
M
M
Rodríguez
C
C
Muñoz
D
D
Fernández de Corres
L
L
D000485:Allergens / Q000187:drug effects / Q000276:immunology / Q000302:isolation & purification*; D000918:Antibody Specificity; D001797:Blood Protein Electrophoresis / Q000379:methods; D003101:Collodion; D017449:Dermatitis, Allergic Contact / Q000097:blood / Q000209:etiology* / Q000276:immunology; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005932:Gloves, Surgical; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D007073:Immunoglobulin E / Q000097:blood / Q000276:immunology*; D007525:Isoelectric Focusing; D008567:Membranes, Artificial; D010940:Plant Proteins / Q000187:drug effects / Q000276:immunology / Q000302:isolation & purification*; D010969:Plastics; D011485:Protein Binding; D011489:Protein Denaturation / Q000187:drug effects; D012408:Rubber / Q000009:adverse effects* / Q000737:chemistry; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology
D000485:Allergens; D008567:Membranes, Artificial; D010940:Plant Proteins; D010969:Plastics; D012967:Sodium Dodecyl Sulfate; D007073:Immunoglobulin E; D003101:Collodion; D012408:Rubber