Development of a hypoallergenic recombinant parvalbumin for first-in-man subcutaneous immunotherapy of fish allergy.
Int Arch Allergy Immunol
; 166(1): 41-51, 2015.
Article
in En
| MEDLINE
| ID: mdl-25765512
ABSTRACT
BACKGROUND:
The FAST (food allergy-specific immunotherapy) project aims at developing safe and effective subcutaneous immunotherapy for fish allergy, using recombinant hypoallergenic carp parvalbumin, Cyp c 1.OBJECTIVES:
Preclinical characterization and good manufacturing practice (GMP) production of mutant Cyp (mCyp) c 1.METHODS:
Escherichia coli-produced mCyp c 1 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 histamine release assay, immunogenicity by immunization of laboratory animals and stimulation of patients' peripheral blood mononuclear cells (PBMCs). Reference molecules were purified wild-type Cyp c 1 (natural and/or recombinant). GMP-compliant alum-adsorbed mCyp c 1 was tested for acute toxicity in mice and rabbits and for repeated-dose toxicity in mice. Accelerated and real-time protocols were used to evaluate stability of mCyp c 1 as drug substance and drug product.RESULTS:
Purified mCyp c 1 behaves as a folded and stable molecule. Using sera of 26 double-blind placebo-controlled food-challenge-proven fish-allergic patients, reduction in allergenic activity ranged from 10- to 5,000-fold (1,000-fold on average), but with retained immunogenicity (immunization in mice/rabbits) and potency to stimulate human PBMCs. Toxicity studies revealed no toxic effects and real-time stability studies on the Al(OH)3-adsorbed drug product demonstrated at least 20 months of stability.CONCLUSION:
The GMP drug product developed for treatment of fish allergy has the characteristics targeted for in FAST i.e. hypoallergenicity with retained immunogenicity. These results have warranted first-in-man immunotherapy studies to evaluate the safety of this innovative vaccine.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Parvalbumins
/
Calcium-Binding Proteins
/
Allergens
/
Desensitization, Immunologic
/
Fish Proteins
/
Food Hypersensitivity
Type of study:
Clinical_trials
/
Guideline
Language:
En
Journal:
Int Arch Allergy Immunol
Journal subject:
ALERGIA E IMUNOLOGIA
Year:
2015
Document type:
Article
Affiliation country:
Países Bajos