ABSTRACT
The potency of allergen extracts is determined as total allergenic activity without consideration of their composition and the units differ from one manufacturer to another, making it very difficult to compare the different products. Recently, purified major allergens have been obtained by recombinant DNA technology and produced under Good Manufacturing Practice (GMP) conditions. In principle, such recombinant allergens could be established as reference standards and could help for the standardisation of the major allergen content of allergen extracts. Two recombinant major allergens, one from birch pollen, rBet v 1, and one from Timothy grass pollen, Phl p 5a, have been selected at the end of the CREATE programme as a potential starting point for the establishment as European Pharmacopoeia (Ph. Eur.) Reference Standards through a project run by the Biological Standardisation Programme (BSP) of the European Directorate for the Quality of Medicines & HealthCare (EDQM). To this end, bulk candidate recombinant materials, produced under GMP conditions, were procured from two European manufacturers and subsequently formulated and lyophilised. Four ELISA systems from three different manufacturers were included in the project, two for Bet v 1 and two for Phl p 5a with the aim of establishing reference methods for determination of the respective major antigens both in natural allergen extracts as well as in recombinant allergen products. The project was run in 3 phases: a preparatory and preliminary testing phase (feasibility phase or Phase 1), an extended feasibility phase carried out in 3 laboratories (Phase 2) to confirm the transferability of the methods and an international collaborative study with a large number of participating laboratories (Phase 3). This article describes the work done in Phase 1 and Phase 2, i.e. the physico-chemical and biological characterisation of the recombinant candidate reference standards, the assessment of their suitability for the intended purpose as well as the evaluation of the candidate ELISA systems. The results show that both candidate reference standards are suitable for the intended purpose. In addition, three out of the four ELISA systems that were included in the preliminary phase were found to be appropriate for further evaluation in the collaborative study which was organised in 2011. The results of the collaborative study will be published separately.
Subject(s)
Allergens/chemistry , Antigens, Plant/chemistry , Enzyme-Linked Immunosorbent Assay/methods , Enzyme-Linked Immunosorbent Assay/standards , Plant Proteins/standards , Pollen/chemistry , Allergens/genetics , Allergens/immunology , Antigens, Plant/genetics , Antigens, Plant/immunology , Basophils/drug effects , Basophils/immunology , Cells, Cultured , Escherichia coli/genetics , Feasibility Studies , Histamine Release/immunology , Humans , Plant Proteins/chemistry , Plant Proteins/genetics , Plant Proteins/immunology , Pollen/genetics , Pollen/immunology , Reproducibility of ResultsSubject(s)
Bacteremia/diagnosis , Bacteremia/immunology , Immunocompromised Host , Mycoplasma Infections/diagnosis , Mycoplasma Infections/immunology , Mycoplasma hominis/isolation & purification , Adult , Anti-Bacterial Agents , Bacteremia/drug therapy , Base Sequence , Diabetes Mellitus, Type 1/diagnosis , Diabetes Mellitus, Type 1/therapy , Disease Progression , Drug Therapy, Combination/administration & dosage , Follow-Up Studies , Humans , Kidney Transplantation/immunology , Male , Molecular Sequence Data , Mycoplasma Infections/drug therapy , Pancreas Transplantation/immunology , Polymerase Chain Reaction , Renal Dialysis , Risk AssessmentABSTRACT
BACKGROUND: Local antimicrobial systems have gained importance, as illustrated by current research on drug delivery systems (DDS). We aimed to develop materials that combine hemostatic and antimicrobial efficacy as well as adhesiveness for use in surgical tissue management. MATERIALS AND METHODS: Materials were evaluated by in vitro studies employing microbiological and technological methods. RESULTS: Antimicrobial impregnation of a collagen fleece, which is a pre-coated fibrinogen-based adhesive and therefore ready-to-use (TachoComb), is significantly more efficient--both in terms of the antimicrobial efficacy (p < 0.001) as well as the adhesive strength (p = 0.03) -than coating an antibiotic-containing collagen fleece "on-site" with fibrin glue. CONCLUSION: Due to ease of practical handling and favorable pharmacoeconomics, this DDS is recommended for both open and minimally invasive surgery.
Subject(s)
Anti-Infective Agents/administration & dosage , Collagen , Drug Delivery Systems/methods , Fibrin Tissue Adhesive , Hemostasis, Surgical/methods , Tissue Adhesives , ImmunodiffusionABSTRACT
A 26-year-old female patient was admitted to the hospital because of bilateral gonarthritis, lymphadenopathy and disseminated chorioiditis as primary manifestation of Lyme borreliosis. Antibody titers against Borrelia burgdorferi did not reach diagnostic levels as determined by an indirect immunofluorescence assay. However, diagnosis was established by the detection of IgG and IgM antibodies in Western blot analysis, and by the demonstration of an enhanced T-cell proliferation to Borrelia burgdorferi in a lymphocyte proliferation assay. This case report indicates that arthritides may already occur in stage II (disseminated infection) of Lyme borreliosis. Therefore, Lyme borreliosis must be considered in patients with chorioiditis and pauciarticular arthritis. In the case of non-diagnostic antibody titers in indirect immunofluorescence tests (or ELISA), Western blot analysis and lymphocyte proliferation assays should be performed in addition.
