Display of the VP1 epitope of foot-and-mouth disease virus on bacteriophage T7 and its application in diagnosis

Foot-and-mouth disease (FMD) is a highly contagious epidemic disease threatening the cattle industry since the sixteenth century. In recent years, the development of diagnostic assays for FMD has benefited considerably from the advances of recombinant DNA technology. In this study, the immunodominant region of the capsid protein VP1 of the foot-and-mouth disease virus (FMDV) was fused to the T7 bacteriophage and expressed on the surface of the bacteriophage capsid protein. The recombinant protein of about 42 kDa was detected by the anti-T7 tag monoclonal antibody in Western blot analysis. Phage ELISA showed that both the vaccinated and positive infected bovine sera reacted significantly with the recombinant T7 particle. This study demonstrated the potential of the T7 phage displaying the VP1 epitope as a diagnostic reagent

Reduction in magnetic coercivity of Co nanomagnets by Fe alloying

We measured the magnetic hysteresis and coercivity of individual Co and Co$_{0.8}$Fe$_{0.2}$ bilayer nano-sized island structures formed on Cu (111) substrate using spin-polarized scanning tunneling microscopy. From the hysteresis taken on various sizes of islands, we found that the alloyed islands are ferromagnetic with out-of-plane magnetic anisotropy, same as the pure islands. Coercivity of the alloy islands, which is dependent on their size, was significantly reduced to ≈40% of that of the pure islands. Based on the Stoner–Wohlfarth model, we evaluated the amount of magnetic anisotropic energy and anisotropy constant for both pure and alloy islands. Since tunneling spectra taken on the alloy islands show upward shifts of the valence electronic states as compared to the pure ones, fewer electrons populated in the valence band of the alloy islands are presumably responsible for the reduction in the magnetic anisotropic energy

The direct recovery of recombinant hepatitis B core antigen from disruptate derived from continuous-flow bead milling

HBcAg (hepatitis B core antigen) is a nanoplex bioproduct that has a great potential in the development of therapeutic drugs and vaccines. In the present study, a continuous-flow bead milling for the disruption of Escherichia coli was optimized and a direct recovery protocol to isolate the recombinant HBcAg from the unclarified E. coli disruptate was developed. The optimal condition for continuous-flow bead milling for the release of HBcAg from E. coli was achieved at a feed flow rate of 15 litres/h, biomass concentration of 10% [ww/v (wet weight/vol.)] and impeller tip speed of 14 m/s. The sucrose-density-gradient analysis showed that the particulate form of the HBcAg released by this optimal condition is still preserved. In the direct purification of HBcAg from the unclarified disruptate, the AE-EBAC (anion-exchange expanded-bed adsorption chromatography) technique was employed. A 54% adsorption and 50.7% recovery of HBcAg were achieved in this direct recovery process. The purity of HBcAg recovered was 49.8%, which corresponds to a purification factor of 2.0. ELISA showed that the HBcAg recovered is functionally active

Multistep Hybrid Iterations for Systems of Generalized Equilibria with Constraints of Several Problems

We first introduce and analyze one multistep iterative algorithm by hybrid shrinking projection method for finding a solution of the system of generalized equilibria with constraints of several problems: the generalized mixed equilibrium problem, finitely many variational inclusions, the minimization problem for a convex and continuously Fréchet differentiable functional, and the fixed-point problem of an asymptotically strict pseudocontractive mapping in the intermediate sense in a real Hilbert space. We prove strong convergence theorem for the iterative algorithm under suitable conditions. On the other hand, we also propose another multistep iterative algorithm involving no shrinking projection method and derive its weak convergence under mild assumptions

Triple Hierarchical Variational Inequalities with Constraints of Mixed Equilibria, Variational Inequalities, Convex Minimization, and Hierarchical Fixed Point Problems

We introduce and analyze a hybrid iterative algorithm by virtue of Korpelevich's extragradient method, viscosity approximation method, hybrid steepest-descent method, and averaged mapping approach to the gradient-projection algorithm. It is proven that under appropriate assumptions, the proposed algorithm converges strongly to a common element of the fixed point set of infinitely many nonexpansive mappings, the solution set of finitely many generalized mixed equilibrium problems (GMEPs), the solution set of finitely many variational inequality problems (VIPs), the solution set of general system of variational inequalities (GSVI), and the set of minimizers of convex minimization problem (CMP), which is just a unique solution of a triple hierarchical variational inequality (THVI) in a real Hilbert space. In addition, we also consider the application of the proposed algorithm to solve a hierarchical fixed point problem with constraints of finitely many GMEPs, finitely many VIPs, GSVI, and CMP. The results obtained in this paper improve and extend the corresponding results announced by many others

Hierarchical Fixed Point Problems in Uniformly Smooth Banach Spaces

We propose some relaxed implicit and explicit viscosity approximation methods for hierarchical fixed point problems for a countable family of nonexpansive mappings in uniformly smooth Banach spaces. These relaxed viscosity approximation methods are based on the well-known viscosity approximation method and hybrid steepest-descent method. We obtain some strong convergence theorems under mild conditions

A preparative hydrophobic interaction chromatography for purification of recombinant nucleocapsid protein of Nipah virus from clarified Escherichia coli homogenate

The downstream processing of the recombinant nucleocapsid (N) protein of Nipah virus (NiV) from Escherichia coli homogenate using a preparative hydrophobic interaction chromatography (HIC) was investigated in the present study. Ammonium sulfate precipitation experiment was performed and it showed that 15% saturation of the salt was the most suitable salt concentration for the binding buffer. Batch binding of the N protein of NiV was performed using Sepharose™ 6 Fast Flow (FF) adsorbents coupling separately with four different types of ligand; phenyl low substitution, phenyl high substitution, butyl and octyl. The phenyl low substitution ligand was selected for subsequent optimization process due to its highest yield and purity of the N protein achieved from the batch binding experiment. The HIC for purification of the N protein of NiV was further scaled-up using a 10 cm column packed with phenyl low substitution Sepharose™ adsorbent. A recovering yield of 81% of the N protein of NiV with a purification factor of 9.3 was achieved from this scaled-up operation. The antigenicity of the purified N protein was still preserved as shown in ELISA analysis

Purification of histidine-tagged nucleocapsid protein of Nipah virus using immobilized metal affinity chromatography

Nucleocapsid (N) protein of Nipah virus (NiV) is a potential serological marker used in the diagnosis of NiV infections. In this study, a rapid and efficient purification system, HisTrapTM 6 Fast Flowpacked bed column was applied to purify recombinant histidine-tagged N protein of NiV from clarified feedstock. The optimizations of binding and elution conditions of N protein of NiV onto and from Nickel SepharoseTM 6 Fast Flow were investigated. The optimal binding was achieved at pH 7.5, superficial velocity of 1.25 cm/min.The bound N protein was successfully recovered by a stepwise elution with different concentration of imidazole (50, 150, 300 and 500 mM). The N protein of NiV was captured and eluted from an inlet N protein concentration of 0.4 mg/ml in a scale-up immobilizedmetal affinity chromatography (IMAC) packed bed column of Nickel SepharoseTM 6 Fast Flow with the optimized condition obtained from the method scouting. The purification of histidine-tagged N protein using IMAC packed bed column has resulted a 68.3% yield and a purification factor of 7.94

A preparative purification process for recombinant Hepatitis B core antigen using online capture by expanded bed adsorption followed by size-exclusion chromatography

Hepatitis B core antigen (HBcAg) is an important serological marker used in the diagnosis of hepatitis B virus (HBV) infections. In the current study, a fast and efficient preparative purification protocol for truncated HBcAg from Escherichia coli disruptate was developed. The recombinant HBcAg was first captured by anion exchange expanded bed adsorption chromatography integrated with a cell disruption process. This online capture process has shortened the process time and eliminated the “hold-up” period that may be detrimental to the quality of target protein. The eluted product from the expanded bed adsorption chromatography was subsequently purified using size-exclusion chromatography. The results showed that this novel purification protocol achieved a recovery yield of 45.1% with a product purity of 88.2%, which corresponds to a purification factor of 4.5. The recovered HBcAg is still biologically active as shown by ELISA test