Purification and immunogenicity of hemagglutinin from highly pathogenic avian influenza virus H5N1 expressed in Nicotiana benthamiana

Highly pathogenic avian influenza (HPAI) H5N1 is an ongoing global health concern due to its severe sporadic outbreaks in Asia, Africa and Europe, which poses a potential pandemic threat. The development of safe and cost-effective vaccine candidates for HPAI is considered the best strategy for managing the disease and addressing the pandemic preparedness. The most potential vaccine candidate is the antigenic determinant of influenza A virus, hemagglutinin (HA). The present research was aimed at developing optimized expression in Nicotiana benthamiana and protein purification process for HA from the Malaysian isolate of H5N1 as a vaccine antigen for HPAI H5N1. Expression of HA from the Malaysian isolate of HPAI in N. benthamiana was confirmed, and more soluble protein was expressed as truncated HA, the HA1 domain over the entire ectodomain of HA. Two different purification processes were evaluated for efficiency in terms of purity and yield. Due to the reduced yield, protein degradation and length of the 3-column purification process, the 2-column method was chosen for target purification. Purified HA1 was found immunogenic in mice inducing H5 HA-specific IgG and a hemagglutination inhibition antibody. This paper offers an alternative production system of a vaccine candidate against a locally circulating HPAI, which has a regional significance

Fator capacidade de fósforo em solos de pernambuco mineralogicamente diferentes e influência do pH na capacidade máxima de adsorção Phosphate capacity factor in mineralogically different soils in Pernambuco and the influence of pH on the maximum capacity of adsorption

O Fator Capacidade de Fósforo (FCP) é definido pela razão de equilíbrio entre o fator quantidade de P (Q) e o fator intensidade (I) e representa uma medida da capacidade do solo em manter um determinado nível de P em solução. As características e o teor dos constituintes minerais da fração argila são responsáveis por uma maior ou menor FCP, interferindo nas relações solo-planta. Por outro lado, o pH do solo tem, em alguns casos, mostrado-se com efeito na adsorção e, em outros, com pequena e não consistente alteração na Capacidade Máxima de Adsorção de P (CMAP). Objetivou-se, neste trabalho, determinar o FCP de solos mineralogicamente diferentes em Pernambuco; correlacionar características físicas e químicas dos solos com o FCP; e avaliar o efeito do pH na CMAP. Amostras subsuperficiais de quatro solos, mineralogicamente diferentes, foram caracterizadas química e fisicamente e determinado o FCP. Essas amostras foram corrigidas com CaCO3 e MgCO3 na proporção 4:1 e incubadas por 30 dias, com exceção do Vertissolo. Determinou-se a CMAP antes e após a correção dos solos. O experimento consistiu de um fatorial 4 x 2 (quatro solos com e sem correção), distribuídos em blocos ao acaso, com três repetições. As características dos solos que melhor refletiram o FCP foram o P remanescente (P-rem) e a CMAP. Independentemente dos constituintes mineralógicos da fração argila, solos com elevados teores de alumínio apresentaram aumento da CMAP com a correção. A energia de adsorção (EA) nos solos corrigidos foi, em média, significativamente menor, independentemente do solo.<br>Phosphate Maximum Capacity (FCP) is defined by the ratio of equilibrium between the amount of factor P (Q) and factor intensity (I) and represents a measure of the soil ability to maintain a certain level of P in solution. The characteristics and content of the constituents of clay minerals are responsible for a greater or lesser FCP, interfering in soil-plant relations. Moreover, the soil pH has affected adsorption, and in other cases, it has shown small and inconsistent change in the maximum adsorption capacity of P (CMAP). Thus, this study aimed to determine the different FCP soil mineralogy in Pernambuco; to correlate physical and chemical characteristics of soils with PBC and to evaluate the effect of pH on the CMAP. Subsurface soil samples from four different soils were characterized chemically and physically determined, and the PBC was determined. These samples were corrected with CaCO3 and MgCO3 in a 4:1 ratio and incubated for 30 days, except the Vertisol. The CMAP was determined before and after correction of the soil. The experiment consisted of a 4 x 2 factorial (four soils with and without correction), distributed in randomized blocks with three replicates. Soil characteristics that best reflected the PBC were the remaining P (P-rem) and MPAC. Regardless of the constituents of clay mineralogy, soil with high aluminum levels had increased CMAP after correction. The energy of adsorption (EA) in the limed soils was on average significantly lower, regardless of the soil