Antigenic analysis (IgE and monoclonal antibodies) of the major shrimp allergen Pen a 1 (Tropomyosin) from Penaeus aztecus.
Pen a 1, the major shrimp allergen from the brown shrimp Penaeus aztecus was purified by preparative SDS-PAGE. Peptides were generated from Pen a 1 by CNBr cleavage and endoproteinase (Lys-C, Glu-C, trypsin, alkaline protease, Arg-C, chymotrypsin) digestion. The molecular weights of the resulting CNBr cleavage and enzymatic digestion products, separated by peptide SDS-PAGE, ranged from 1.5 to 20 kD. Following SDS-PAGE and semidry blotting, the analysis of monoclonal antibody (mAb) and subjects' IgE reactivities demonstrated that with the exception of alkaline protease, all cleavage procedures yielded IgE-binding peptides. However, since not all peptides of every digest bind IgE, it appears that IgE-binding epitopes are restricted to certain parts of the Pen a 1 molecule. mAbs bound to CNBr, Lys-C, trypsin, Glu-C and Arg-C peptides. Since mAbs reacted to several peptides from the same digest, Pen a 1 may have several similar epitopes. The comparison of IgE and mAb reactivities demonstrated similar but not identical binding patterns.
Reese
G
G
Section of Clinical Immunology and Allergy, Tulane University Medical Center, New Orleans, LA 70112, USA.
Daul
C B
CB
Lehrer
S B
SB
D000485:Allergens / Q000737:chemistry / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D060829:Arthropod Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D007073:Immunoglobulin E / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D033561:Penaeidae / Q000276:immunology*; D010449:Peptide Mapping; D011506:Proteins / Q000737:chemistry / Q000276:immunology*; D014335:Tropomyosin / Q000737:chemistry / Q000276:immunology*
D000485:Allergens; D000911:Antibodies, Monoclonal; D060829:Arthropod Proteins; D000939:Epitopes; D011506:Proteins; D014335:Tropomyosin; C058449:shrimp allergens Sa; D007073:Immunoglobulin E