Gain of structure and IgE epitopes by eukaryotic expression of the major Timothy grass pollen allergen, Phl p 1.

Approximately 400 million allergic patients are sensitized against group 1 grass pollen allergens, a family of highly cross-reactive allergens present in all grass species. We report the eukaryotic expression of the group 1 allergen from Timothy grass, Phl p 1, in baculovirus-infected insect cells. Domain elucidation by limited proteolysis and mass spectrometry of the purified recombinant glycoprotein indicates that the C-terminal 40% of Phl p 1, a major IgE-reactive segment, represents a stable domain.

Recombinant protein to analyze autoantibodies to proteinase 3 in systemic vasculitis.

The presence of antineutrophil cytoplasmic autoantibodies with specificity for proteinase 3 (PR3-ANCA) usually is detected by enzyme-linked immunosorbent assay (ELISA) with purified PR3 as a substrate. We studied the technical performance of direct and capture ELISA using a recombinant proteolytically inactive form of PR3 produced in the baculovirus expression system for the detection of PR3-ANCA in 114 patients with systemic vasculitis at diagnosis. We found that ELISA using recombinant PR3 produced in insect cells is a promising alternative for ELISA with native PR3.

Recombinant autoantigens for diagnosis and therapy of autoimmune diseases.

The characteristic of autoimmune diseases resides in the existence of T- and B-cell autoreactivity when directed against self-proteins. Thus autoantibodies are considered to be of diagnostic relevance. The advent of recombinant protein expression technology has made it possible to produce economically high-quality autoantigens for use in immunoassays for detecting autoantibodies in patients' sera.