Search Results (4)

Background/Objectives: An assay for protein content is essential but insufficient for quality control of acellular pertussis vaccines, which might consist of up to five components, each needing individual quantification. Generally, purified pertussis antigens such as pertussis toxin (PTx), filamentous haemagglutinin (FHA), and pertactin (PRN) should be detoxified or stabilized chemically before being formulated into vaccine bulk. The use of chemical agents like formaldehyde and glutaraldehyde can alter the immunological reactivity of these antigens, rendering direct assays by methods such as ELISA ineffective. Methods: In this study, a simple method based on single radial diffusion (SRD) using low concentrations of polyclonal antisera against PT toxoid (PTd), FHA, and PRN was developed. By adding a detergent, diffusible subunits are produced regardless of the original physical state of the antigens, making it suitable for quantifying these antigens after chemical treatment. Results: The assay has shown good specificity, accuracy, and precision. Furthermore, it can differentiate between preparations with the same protein concentration but different antigenic contents. A significant positive correlation between the antigen content and the in vivo immunogenicity has also been demonstrated. Conclusions: An assay for quality control and consistency monitoring of combined vaccines containing acellular pertussis antigen components has been established. Full article

Assaying the potency of inactivated viral influenza vaccines is performed using single radial immunodiffusion, which is the globally accepted release method for potency. Under conditions of a rapidly emerging pandemic, such as the 2009 H1N1 influenza pandemic, a recognized obstacle in the delivery of vaccines to the public is the time needed for the distribution of calibrated SRID reagents (antisera and antigen standards) to vaccine manufacturers. Previously, we first described a novel streamlined MS-based assay, CombE-IDMS, which does not rely on antisera/antibodies or reference antigens, as a potential rapidly deployable alternate potency method through a comparison with SRID on adjuvanted seasonal quadrivalent vaccine cell-based (aQIVc) materials. In this report, we further demonstrate that the CombE-IDMS method can also be applied to measure the potency of pre-pandemic H5N1 and H5N8 monovalent vaccine materials, each subtype both unadjuvanted and adjuvanted, through a forced degradation study. Overall, CombE-IDMS results align with those of the gold standard SRID method on both H5N1 and H5N8 materials under conditions of thermal, pH, oxidative and freeze/thaw stress, lending further evidence for the CombE-IDMS method’s suitability as an alternate assay for potency of both seasonal and pandemic influenza vaccines. Full article

The aim of the present study was to evaluate the effect of two types of nutritional supplementation during late gestation on the chemical composition, energy value, and IgG concentration in the colostrum and the IgG concentration in the blood serum of lambs. Pregnant Merino Precoz ewes (n = 36) carrying single fetuses were used. Animals were kept grazing on the Mediterranean annual grassland. From day ~90 of pregnancy, animals were allocated into three groups: daily supplementation with oat grain or lupine grain and a control group without supplementation. Immediately after parturition, colostrum was collected from each ewe, and a blood sample was taken from the lambs 24 h after birth. For the evaluation of the chemical composition of the colostrum, an EKOMILK^® milk analyzer was used. The energy value of the colostrum was calorimetrically evaluated. IgG concentrations were measured by simple radial immunodiffusion. Data were analyzed by analysis of variance. Colostrum content of protein and non-fat solids was higher in the group supplemented with oat grain than in the lupine grain supplemented and control groups (p ≤ 0.05). In contrast, ewes supplemented with lupine grain had the highest concentration of fat in their colostrum (p ≤ 0.05). Oat grain supplementation resulted in higher concentrations of IgG, both in sheep colostrum and in the blood serum of their lambs (p ≤ 0.05), being higher than those observed in the lupine grain and control groups. Ewes that gave birth to male lambs had significantly higher concentrations of IgG in their colostrum compared to ewes that gave birth to females (p ≤ 0.05). The colostral IgG concentration positively correlated with the serum IgG concentration of the lambs (r = 0.32; p ≤ 0.05). The results indicate that the quality of colostrum and the immunological status of the newborn lambs can be improved by supplementation with oat grain. Full article

Intravenous iron preparations are typically classified as non-dextran-based or dextran/dextran-based complexes. The carbohydrate shell for each of these preparations is unique and is key in determining the various physicochemical properties, the metabolic pathway, and the immunogenicity of the iron-carbohydrate complex. As intravenous dextran can cause severe, antibody-mediated dextran-induced anaphylactic reactions (DIAR), the purpose of this study was to explore the potential of various intravenous iron preparations, non-dextran-based or dextran/dextran-based, to induce these reactions. An IgG-isotype mouse monoclonal anti-dextran antibody (5E7H3) and an enzyme-linked immunosorbent assay (ELISA) were developed to investigate the dextran antigenicity of low molecular weight iron dextran, ferumoxytol, iron isomaltoside 1000, ferric gluconate, iron sucrose and ferric carboxymaltose, as well as isomaltoside 1000, the isolated carbohydrate component of iron isomaltoside 1000. Low molecular weight iron dextran, as well as dextran-based ferumoxytol and iron isomaltoside 1000, reacted with 5E7H3, whereas ferric carboxymaltose, iron sucrose, sodium ferric gluconate, and isolated isomaltoside 1000 did not. Consistent results were obtained with reverse single radial immunodiffusion assay. The results strongly support the hypothesis that, while the carbohydrate alone (isomaltoside 1000) does not form immune complexes with anti-dextran antibodies, iron isomaltoside 1000 complex reacts with anti-dextran antibodies by forming multivalent immune complexes. Moreover, non-dextran based preparations, such as iron sucrose and ferric carboxymaltose, do not react with anti-dextran antibodies. This assay allows to assess the theoretical possibility of a substance to induce antibody-mediated DIARs. Nevertheless, as this is only one possible mechanism that may cause a hypersensitivity reaction, a broader set of assays will be required to get an understanding of the mechanisms that may lead to intravenous iron-induced hypersensitivity reactions. Full article