Development of Monoclonal Mouse Antibodies Specific for Allergenic Components in Ryegrass <i>(Lolium perenne)</i> Pollen

Abstract

The development of monoclonal mouse antibodies against ryegrass (Lolium perenne) pollen allergens is described. Hybridoma colonies secreting antibodies specific for allergenic components were detected using an enzyme-linked immunosorbent assay. Positive colonies were cloned and expanded. The pollen components with which the monoclonal antibodies interact were identified and characterised following sodium dodecyl sulphate polyacrylamide gel electrophoresis and electrophoretic transfer to nitrocellulose. In this paper six monoclonal mouse antibodies are described. Three antibodies interact with a single molecule of between 30,000 and 35,000 daltons. One antibody interacts with a component of 16,000 daltons whereas the remaining two antibodies react with more than one component, one reacting with two components at 28,000 and 30,000, and the other with five components having molecular weights between 18,000 and 71,000 daltons.

Isolation of cDNA encoding a newly identified major allergenic protein of rye-grass pollen: intracellular targeting to the amyloplast.

“…Conditions for electrophoresis and immunoblotting were essentially as described (12). Monoclonal antibodies (mAbs) 3.2 and 21.3 bind specifically to the Lol pI group of antigens (9); mAb 40.1, although originating in a different laboratory, shows similar specificity (12,15). mAb 12.3 binds to a 31-kDa component in the Lol pI group (15).…”

“…Monoclonal antibodies (mAbs) 3.2 and 21.3 bind specifically to the Lol pI group of antigens (9); mAb 40.1, although originating in a different laboratory, shows similar specificity (12,15). mAb 12.3 binds to a 31-kDa component in the Lol pI group (15). For IgE antibody binding, blots were incubated in allergic sera collected from at least four patients with high skin sensitivity for grass pollen, pooled and used diluted 1: 5 in Tris-buffered saline (TBS: 150 mM NaCI/10 mM Tris-HCI, pH 7.5) containing 0.5% bovine serum albumin.…”

Standardization of Allergens

“…1, part 2, Jan. 1983). The methods employed by Smart et a1 (64) should prove generally applicable to a variety of unfractionated allergen sources. ELISA was used both to screen hybridoma supernatants and for the detection of IgE-binding activity while specificities of monoclonal antibodies were examined by reaction with ryegrass pollen antigens following electrophoretic transfer to nitrocellulose sheets.…”

Molecular cloning and immunological characterisation of Cyn d 7, a novel calcium‐binding allergen from Bermuda grass pollen1

“…The major outdoor cause of allergy is airborne grass pollen [2]. In warm temperate and sub-tropical climates, such as northern and central Australia, Taiwan, southwestern United States and South Africa, Bermuda grass {Cynodon dactylon), is an important source of allergens.…”