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Recombinant allergen Lol p II: expression, purification and characterization
E Tamborini1, A Brandazza, C De Lalla
1Department of Biology and Technology, San Raffaele Scientific Institute, Milano, Italy.
Molecular Immunology
|May 1, 1995
Summary
Researchers produced a recombinant version of the major rye grass allergen, Lol p II, in E. coli. This non-fusion protein is stable, soluble, and immunologically similar to the natural allergen, showing potential for allergy research.
Area of Science:
- Allergen research
- Molecular biology
- Immunology
Background:
- Perennial rye grass (Lolium perenne) pollen is a significant global cause of Type I allergies.
- Lol p II is identified as a major allergen within rye grass pollen.
- Previous work involved cloning, sequencing, and expressing Lol p II fused to human ferritin.
Purpose of the Study:
- To describe the expression, purification, and characterization of recombinant Lol p II.
- To produce Lol p II as a non-fusion protein in E. coli.
- To assess the immunological and biochemical properties of the recombinant allergen.
Main Methods:
- Expression of recombinant Lol p II in the periplasm of E. coli.
- Purification of the non-fusion recombinant protein.
- Immunological assays including IgE binding competition and skin prick tests.
- Biochemical analyses of stability, solubility, and molecular behavior.
Main Results:
- High yields of recombinant Lol p II were achieved.
- The purified protein was easily obtained in milligram quantities.
- Recombinant Lol p II competed with natural Lol p II for specific IgE binding.
- Skin prick tests confirmed the induction of allergic responses, indicating immunological analogy to the natural protein.
- Biochemical analysis revealed recombinant Lol p II as a stable, soluble, monomeric, globular protein.
Conclusions:
- Recombinant Lol p II can be successfully overexpressed and purified from E. coli.
- The recombinant allergen is immunologically equivalent to the natural Lol p II.
- The characterized properties suggest its utility in further allergy studies and potential therapeutic strategies.