Analysis of ragweed allergens recognized by monoclonal antibody 8-5 using two-dimensional gel electrophoresis and immunoblotting.
A series of MoAbs against major allergens (Amb a I and Amb a II) of the short ragweed pollen is reported by this laboratory. One of these MoAbs, 8-5, showed strong reactivity to Amb a II, weak reactivity to Amb a I-B and I-C, and no reactivity to Amb a I-B2 of the short ragweed pollen by SDS-PAGE and immunoblotting. In the present study, ragweed allergens recognized by MoAb 8-5 were further analyzed by two-dimensional gel electrophoresis and immunoblotting. The results showed that MoAb 36-6, which has been identified to be reactive with Amb a I only, reacted mainly with components with MW of about 39 kD, which corresponded to creatine kinase charge standards from -14 to -20 in both the Amb a I-B and I-C preparation and the crude extract of the short ragweed pollen. However, MoAb 8-5 showed strong reactivity to components with about the same MW but charge standards from -8 to -13 in Amb a II and the crude extract of the short ragweed pollen. Furthermore, a weak positive reactivity of MoAb 8-5 to components with MW of about 39 kD and charge standards from -8 to -14 in Amb a I-B and I-C was also observed. It was thus concluded that MoAb 8-5 reacted with Amb a II only.(ABSTRACT TRUNCATED AT 250 WORDS)
Shen
H D
HD
Department of Medical Research, Veterans General Hospital-Taipei, Taiwan, Republic of China.
Lin
W L
WL
Han
S H
SH
D000485:Allergens / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D052179:Antigens, Plant; D015180:Electrophoresis, Gel, Two-Dimensional; D004797:Enzyme-Linked Immunosorbent Assay; D015151:Immunoblotting; D010940:Plant Proteins / Q000276:immunology*; D011058:Pollen / Q000276:immunology*
D000485:Allergens; C026967:Amb a I protein, Ambrosia artemisiifolia; D000911:Antibodies, Monoclonal; D052179:Antigens, Plant; D010940:Plant Proteins; C074244:Amb a II protein, Ambrosia artemisiifolia