5 research outputs found
Ionometric sensor for ammonium ions control in sewage waters
Цель работы заключалась в создании измерительного датчика для автоматизированного потенциометрического контроля содержания ионов аммония в проточных условиях. В результате исследования разработана конструкция электрода сравнения измерительного датчика, представляющая собой ионоселективный электрод, опущенный в буферную систему с катионитом. В работе представлен потенциометрический анализ модельных растворов ионов аммония, изучено мешающее влияние температуры, а также определена стабильность предложенной системы. The purpose of the work was to create a sensor for automated potentiometric monitoring of the ammonium ions content in flowing conditions. As a result of the study, the reference electrode design was developed. This is an ion-selective electrode lowered into a buffer system with cation exchanger. The work presents a potentiometric analysis of ammonium ions model solutions, the study of temperature interfering effect, and system's stability determination
Development of a hypoallergenic recombinant parvalbumin for first-in-man subcutaneous immunotherapy of fish allergy
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. © 2015 S. Karger AG, Basel
Nuclear Magnetic Resonance (NMR) Solution Structure, Dynamics, and Binding Properties of the Kringle IV Type 8 Module of Apolipoprotein(a)
Supplementary Material for: Development of a Hypoallergenic Recombinant Parvalbumin for First-in-Man Subcutaneous Immunotherapy of Fish Allergy
<p><b><i>Background:</i></b> 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. <b><i>Objectives: </i></b>Preclinical characterization and good manufacturing practice (GMP) production of mutant Cyp (mCyp) c 1. <b><i>Methods:</i></b><i>Escherichia coli</i>-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. <b><i>Results:</i></b> 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)<sub>3</sub>-adsorbed drug product demonstrated at least 20 months of stability. <b><i>Conclusion:</i></b>
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.</p