Development of a Hypoallergenic Recombinant XXXX for First-in- @entity1 Subcutaneous Immunotherapy of Fish Allergy.

@entity1 @entity137 @entity9963 @entity6746 @entity19 @entity566 @entity95 @entity112 @entity12525 @entity13323 @entity7136 @entity15537 @entity526
BACKGROUND: The FAST (food @entity566 -specific immunotherapy) project aims at developing safe and effective subcutaneous immunotherapy for @entity9963 , using recombinant hypoallergenic @entity7136 @entity15537 , @entity13323 @entity6746 . OBJECTIVES: Preclinical characterization and good manufacturing practice (GMP) production of mutant @entity13323 (mCyp) @entity6746 . METHODS: @entity526 -produced mCyp @entity6746 was purified using standard chromatographic techniques. Physicochemical properties were investigated by gel electrophoresis, size exclusion chromatography, circular dichroism spectroscopy, reverse-phase high-performance liquid chromatography and mass spectrometry. Allergenicity was assessed by ImmunoCAP inhibition and basophil @entity112 release assay, immunogenicity by immunization of laboratory animals and stimulation of @entity1 ' peripheral blood mononuclear cells (PBMCs). Reference molecules were purified wild-type @entity13323 @entity6746 (natural and/or recombinant). GMP-compliant alum-adsorbed mCyp @entity6746 was tested for acute @entity137 in @entity19 and @entity95 and for repeated-dose @entity137 in @entity19 . Accelerated and real-time protocols were used to evaluate stability of mCyp @entity6746 as drug substance and drug product. RESULTS: Purified mCyp @entity6746 behaves as a folded and stable molecule. Using sera of 26 double-blind placebo-controlled food-challenge-proven fish-allergic @entity1 , reduction in allergenic activity ranged from 10- to 5,000-fold (1,000-fold on average), but with retained immunogenicity (immunization in @entity19 / @entity95 ) and potency to stimulate @entity1 PBMCs. @entity137 studies revealed no toxic effects and real-time stability studies on the @entity12525 -adsorbed drug product demonstrated at least 20 months of stability. CONCLUSION: The GMP drug product developed for treatment of @entity9963 has the characteristics targeted for in FAST: i.e. hypoallergenicity with retained immunogenicity. These results have warranted first-in- @entity1 immunotherapy studies to evaluate the safety of this innovative vaccine.    2015 S. Karger AG, Basel.
