Search

Introduction. It is estimated that 50% of vaccines produced annu- ally are wasted because effectivity is dependent on protein structure and heat exposure disrupts the intermolecular interactions that maintain this structure. Since 90% of vaccines require a temperature- controlled supply chain, it is necessary to create a cold chain system to minimize vaccine waste. We have developed a more sustainable technology via the adsorption of Invasion Plasmid Antigen D (IpaD) onto mesoporous silica gels, improving the thermal stability of pro- tein-based therapeutics. Methods.xThe solution depletion method using UV-Vis was uti- lized to study the adsorption of IpaD onto silica gels. The silica-IpaD complex is heated above the denaturing temperature of the protein and then the IpaD is removed using N,N-Dimethyldodecylamine N-oxide (LDAO) and their secondary structure is tested using cir- cular dichroism (CD). Results. Pore diameter, pore volume and surface area were charac- terized for seven different silica gels. Silica gels designated as 6389, 6378, and 6375 had an adsorption percentage above 95% at pore volumes of 2.2, 2.8 and 3.8 cm3 mg-1, respectively. CD analyses con- firmed that the adsorbed IpaD after the heat treatment displayed a similar “W” shape CD signal as the native IpaD, indicating the con- servation of α-helices. In contrast, the unprotected IpaD after being exposed to high temperature shows a flat CD signal, demonstrating the loss of secondary structure. Conclusion. We have successfully increased the thermo-tolerance for IpaD using mesoporous silica and continue to further optimize mesoporous silica’s physiochemical properties to improve adsorption and desorption yields.

Approximately half of all vaccines produced annually are wasted because effectivity is dependent on protein structure and heat exposure disrupts the intermolecular interactions needed to maintain the structure. Thus, most vaccines require a temperature-controlled supply chain to minimize waste. A more sustainable technology was developed via the adsorption of invasion plasmid antigen D (IpaD) onto mesoporous silica, improving the thermal stability of this protein-based therapeutic. Seven silicas were characterized to determine the effects of pore diameter, pore volume, and surface area on protein adsorption. The silica-IpaD complex was then heated above the IpaD denaturing temperature and N,N-dimethyldodecylamine N-oxide was used to remove IpaD from the silica. Circular dichroism confirmed that the adsorbed IpaD after the heat treatment maintained a native secondary structure rich in α-helix content. In contrast, the unprotected IpaD after heat treatment lost its secondary structure. Isotherms using Langmuir, Freundlich, and Temkin models demonstrated that the adsorption of IpaD onto silicas is best fit by the Langmuir model. If pores are less than 15 nm, adsorption is negligible. If the pores are between 15 and 25 nm, then monolayer coverage is achieved and IpaD is protected from thermal denaturing. If pores are larger than 25 nm, the adsorption is a multilayer coverage and it is easier to remove the protein from the silica because of a less-developed hydrogen bond network. This case study provides strong evidence that IpaD is thermally stabilized via adsorption on mesoporous silica with the proper range of pore sizes.

Three N-ethyl­carbazole derivatives have been synthesized and tested for non-linear optical (NLO) properties. The compounds are 2-(9-ethyl-9H-carbazol-3-yl­methyl­ene)­malono­nitrile, C18H13N3, (IIIa), 2-cyano-3-(9-ethyl-9H-carba­zol-3-yl)­thio­acryl­amide, C18H15N3S, (IIIb), and 3-(9-ethyl-9H-carbazol-3-yl)-2-(4-phenyl-1,3-thia­zol-2-yl)­acrylo­nitrile, C26H19N3S, (V). It was found that the mol­ecules of (IIIa) and (V) are nearly planar, while non-planarity is more pronounced for (IIIb). Molecules of (IIIa) and (V) exhibit herring-bone packing motifs. In (IIIb), the mol­ecules form layers coplanar with (\overline 201), within which they form centrosymmetric dimers via N—H⋯S hydrogen bonds.