Development and Application of Synthetic Affinity Ligands for the Purification of Ferritin-Based Influenza Antigens

A recently developed novel recombinant influenza antigen vaccine has shown great success in preclinical studies in ferrets and mice. It provides broader protection, and is efficient to manufacture compared to the conventional trivalent influenza vaccines (TIV). Each strain of the recombinant antigen has a constant self-assembled bacterial ferritin core which, if used as a target for affinity chromatography, could lead to a universal purification method. Ferritin in silico models were used to explore potential target binding sites against ligands synthesized by the four-component Ugi reaction. Two ligands, SJ047 and SJ055, were synthesized in solution, characterized by 1H, 13C, and 2D NMR spectroscopy, and subsequently immobilized on the PEG-functionalized beads. Ligands SJ047 and SJ055 displayed apparent Kd values of 2.04 × 10–7 M and 1.91 × 10–8 M, respectively, against the ferritin. SJ047 and SJ055-functionalized resins were able to purify hemagglutinin (New Caledonia)-ferritin expressed in a crude Human Embryonic Kidney (HEK) cell supernatant in a single step to a purity of 85 ± 0.5% (97 ± 1% yield) and 87.5 ± 0.5% (95.5 ± 1.5% yield), respectively. Additionally, SJ047 and SJ055-functionalized resins purified the recombinant antigens when spiked at known concentrations into HEK supernatants. All three strains, hemagglutinin (New Caledonia)-ferritin, hemagglutinin (California)-ferritin, and hemagglutinin (Singapore)-ferritin were purified, thereby offering an ideal alternate platform for affinity chromatography. Following elution from the affinity adsorbents, absorbance at 350 nm showed that there was no aggregation of the recombinant antigens and dynamic light scattering studies further confirmed the structural integrity of the recombinant antigen. The use of Ugi ligands coupled to a PEG-spacer arm to target the ferritin core of the strain is entirely novel and provides an efficient purification of these recombinant antigens. This approach represents a potentially universal method to purify any ferritin-based vaccine

Thermodynamically self-assembling porphyrin-stoppered rotaxanes

A variety of porphyrin-stoppered rotaxanes has been assembled under conditions of thermodynamic reversibility by judicious choice of components and temperatures. An admixture of a thread unit comprising a central naphthodiimide with terminal pyridines (2a), a ring unit dinaphtho-38-crown-10 (1) and either ZnII (3a), RuII (CO) (3b) or RhIII I (3c) as stoppers was shown to form an equilibrating mixture of pseudorotaxanes and the porphyrin-stoppered rotaxanes. The intact rotaxane could be crystallised from solution and chromatographed at low temperatures; at higher temperatures only a mixture of the components separated on chromatography. 1H NMR revealed NOE correlations between all three components for the rotaxane, and exchange peaks for related free and complexed species

Host-guest interactions in acid-porphyrin complexes.

In this report we use the weak interactions of acid-porphyrin complexes to selectively bind competing acids to the faces of a rigid cyclic porphyrin dimer, and characterise the resulting interactions by NMR spectroscopy and nano-electrospray ionisation spectrometry

A self-assembling polymer-bound rotaxane under thermodynamic control

The thermodynamically controlled self-assembly of a neutral donor–acceptor rotaxane, stoppered via porphyrin coordination and bound to polystyrene beads is described, and the dynamic equilibrium between solid and solution phases has been examined by HR MAS nmr spectroscopy

Gel-phase HR-MAS ¹H NMR spectroscopy as a probe for solid-tethered diimide rotaxanes and catenanes

The design and kinetically-controlled construction of a series of solid-tethered supramolecular systems utilising crown ether–naphthalene diimide host–guest chemistry are described.Functionalised polystyrene beads (ArgoGel-OH™) were utilised as the gel-phase solid support for the assembly of an appended diimide-crown catenane, and for a porphyrin-stoppered diimidecrown rotaxane. The structures of the resulting solid-tethered systems were probed using gelphase high-resolution magic-angle spinning (HR-MAS) ¹H NMR spectroscopy. The advantages of using this spectroscopic tool in conjunction with optical microscopy to probe solid-tethered supramolecular systems are discussed

Overview of Natural Gas Boiler Optimization Technologies and Potential Applications on Gas Load Balancing Services

Natural gas is a fossil fuel that has been widely used for various purposes, including residential and industrial applications. The combustion of natural gas, despite being more environmentally friendly than other fossil fuels such as petroleum, yields significant amounts of greenhouse gas emissions. Therefore, the optimization of natural gas consumption is a vital process in order to ensure that emission targets are met worldwide. Regarding residential consumption, advancements in terms of boiler technology, such as the usage of condensing boilers, have played a significant role in moving towards this direction. On top of that, the emergence of technologies such as smart homes, Internet of Things, and artificial intelligence provides opportunities for the development of automated optimization solutions, which can utilize data acquired from the boiler and various sensors in real-time, implement consumption forecasting methodologies, and accordingly provide control instructions in order to ensure optimal boiler functionality. Apart from energy consumption minimization, manual and automated optimization solutions can be utilized for balancing purposes, including natural gas demand response, which has not been sufficiently covered in the existing literature, despite its potential for the gas balancing market. Despite the existence of few research works and solutions regarding pure gas DR, the concept of an integrated demand response has been more widely researched, with the existing literature displaying promising results from the co-optimization of natural gas along with other energy sources, such as electricity and heat

Overview of Natural Gas Boiler Optimization Technologies and Potential Applications on Gas Load Balancing Services

Natural gas is a fossil fuel that has been widely used for various purposes, including residential and industrial applications. The combustion of natural gas, despite being more environmentally friendly than other fossil fuels such as petroleum, yields significant amounts of greenhouse gas emissions. Therefore, the optimization of natural gas consumption is a vital process in order to ensure that emission targets are met worldwide. Regarding residential consumption, advancements in terms of boiler technology, such as the usage of condensing boilers, have played a significant role in moving towards this direction. On top of that, the emergence of technologies such as smart homes, Internet of Things, and artificial intelligence provides opportunities for the development of automated optimization solutions, which can utilize data acquired from the boiler and various sensors in real-time, implement consumption forecasting methodologies, and accordingly provide control instructions in order to ensure optimal boiler functionality. Apart from energy consumption minimization, manual and automated optimization solutions can be utilized for balancing purposes, including natural gas demand response, which has not been sufficiently covered in the existing literature, despite its potential for the gas balancing market. Despite the existence of few research works and solutions regarding pure gas DR, the concept of an integrated demand response has been more widely researched, with the existing literature displaying promising results from the co-optimization of natural gas along with other energy sources, such as electricity and heat