Affinity resins enable a multi-vaccine/platform purification process and rapid response to new viruses

Established platform processes applicable to a range of recombinant sub-unit vaccines and VLPs will enable rapid response as well as easy adaption to seasonal variants. Affinity purification has the ability to provide a platform purification process affording a standard high purity, high yield purification step towards different vaccines. We describe several affinity resins that we have developed that enables this platform approach for different vaccine modalities, some of which can be considered custom affinity resins, whilst the others open access catalog resins. The approach used is based on targeting elements or domains that are common to a vaccine technology and generating affinity resin towards these domains. For example, many vaccines are based upon trimeric virus fusion proteins and incorporate a trimerization domain to ensure correct folding and presentation. By targeting the trimerization domain a platform affinity resin can be developed that yields a platform production process irrespective of which virus protein is fused to the domains. This purification approach can be applied to many trimerization domains irrespective if the domain is proprietary and or with freedom to operate and in public use, and to other domains encourage the formation of larger protein particles (nanoparticles) to elicit immune responses. We present a custom affinity resin towards a proprietary domain that has wide applicability to many sub-unit vaccines within a company’s pipeline. We also present an open access resin that is under development that targets the T4 foldon trimerization domain. The foldon domain is the most widely used trimerization domain in academia and industry and has been used in a multitude of protein subunit vaccines. We further extend this catalog resin platform approach towards whole virus particles for which we have developed an affinity resin that binds to the phospholipid on the outer surface of the lipid membrane. It is our hope and also the intention of the companies that we collaborate with, that these resins will enable producers of these traditional vaccine modalities to effect rapid process development and enable them access to the timescale of development exhibited by mRNA vaccines

A structural analysis of heparin-like glycosaminoglycans using MALDI-TOF mass spectrometry

Abstract. Mass spectrometry (MS) techniques have spear-headed the field of proteomics. Recently, MS has been used to structurally analyse carbohydrates. The heparin/heparan sulfate-like glycosaminoglycans (HLGAGs) present a special set of difficulties for structural analysis because they are highly sulfated and heterogeneous. We have used a matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-MS) technique in which heparin fragments are non-covalently bound to basic peptides of a known mass, so as to limit in-source desulfation and hence afford an accurate mass. We examined a range of different sized fragments with varying degrees of sulfation. The potential of combining the MALDI-MS technique with enzymatic digestion to obtain saccharide sequence information on heparin fragments was explored. A disaccharide analysis greatly assists in determining a sequence from MALDI-MS data. Enzymatic digestion followed by MALDI-MS allows structural data on heparin fragments too large for direct MALDI-MS to be obtained. We demonstrate that synthetic sulfated oligosaccharides can also be analysed by MALDI-MS. There are advantages and limitations with this methodology, but until superior MS techniques become readily accessible to biomedical scientists the MALDI-MS method provides a means to structurally analyse HLGAG fragments that have therapeutic potential because of their ability to bind to and functionally regulate a host of clinically important proteins

The reaction of acetylacetone with amino sugars: implications for the formation of glycosylpyrazole derivatives

Glycosylpyrazoles are efficiently formed by reaction of saccharide hydrazones with pentan-2,4-dione (acetylacetone), but in aqueous buffer, pyrazole derivatives of amino sugars couple with a further equivalent of acetylacetone affording high yields of ketoenamines. These ketoenamines were considerably more stable than the ketoenamines formed from 2-amino-2-deoxy aldoses that have been described as intermediates in the classical Elson-Morgan reaction. Moreover, high yields of perketoenamine derivatives were achieved with oligosaccharides derived from hydrolysis of chitosan. The removal of the ketoenamine moieties to regenerate the free amine was readily accomplished with aqueous hydrazine.8 page(s

Herapin mimetics

Explorations of the therapeutic potential of heparin mimetics, anionic compounds that are analogues of glycosaminoglycans (GAGs), have gone hand-in-hand with the emergence of understanding as to the role of GAGs in many essential biological processes. A myriad of structurally different heparin mimetics have been prepared and examined in many diverse applications. They range in complexity from heterogeneous polysaccharides that have been chemically sulphated to well-defined compounds, designed in part to mimic the natural ligand, but with binding specificity and potency increased by conjugation to non-carbohydrate pharmacophores. The maturity of the field is illustrated by the seven heparin mimetics that have achieved marketing approval and there are several more in late-stage clinical development. An overview of the structural determinants of heparin mimetics is presented together with an indication of their activities. The challenges in developing heparin mimetics as drugs, specificity and potential toxicity issues, are highlighted. Finally, the development path of three structurally very different mimetics, PI-88®, GMI-1070 and RGTAs, each of which is in clinical trials, is described

Heterocyclic derivatives of sugars: the formation of 1-glycosyl-3-methylpyrazol-5-ones from hydrazones

9 page(s

Is it true? (When) does it matter? The roles of likelihood and desirability in argument judgments and attitudes

Several theoretical perspectives either directly or indirectly specify roles for likelihood and desirability information in argument judgments and attitude formation. Some perspectives assume that argument judgments and attitudes are a function of the likelihood of the consequences or conclusions, others contend that the desirability of the consequences or conclusions underlie judgments and attitudes, and expectancy-value perspectives, (e.g., Fishbein, 1963) propose that judgments and attitudes should depend on the likelihood × desirability interaction. Construal level theory (CLT; Trope & Liberman, 2003) also suggests that both likelihood and desirability information impact argument judgments and attitudes, but the roles of each are moderated by when the outcomes are to occur. Three studies examined the sometimes-competing predictions regarding the roles of these variables by orthogonally manipulating levels of likelihood and desirability. Although likelihood and desirability both emerged as components of argument strength, and contributed to attitudes, all 3 studies showed that desirability information was more closely associated with argument strength and attitudes than was likelihood information. In Study 1a, argument strength was shown to mediate the desirability-attitude relation. The likelihood × desirability interaction did not predict attitudes in a manner consistent with expectancy-value predictions, though in some instances likelihood and desirability judgments interacted to predict attitudes and attitude change in the predicted expectancy-value pattern. Studies 1b and 2 showed that the desirability-attitude relation was best described as a cubic trend consistent with prospect theory. CLT predictions examined in Study 2 were largely unsupported. Theoretical and methodological implications are discussed.