The composition and structural properties of lignocellulosic biomass have significant effects on its downstream conversion to fuels, biomaterials, and building-block chemicals. Specifically, the recalcitrance to modification and compositional variability of lignocellulose make it challenging to optimize and control the conditions under which the conversion takes place. Various characterization protocols have been developed over the past 150 years to elucidate the structural properties and compositional patterns that affect the processing of lignocellulose. Early characterization techniques were developed to estimate the relative digestibility and nutritional value of plant material after ingestion by ruminants and humans alike (e.g. dietary fiber). Over the years, these empirical techniques have evolved into statistical approaches that give a broader and more informative analysis of lignocellulose for conversion processes, to the point where an entire compositional and structural analysis of lignocellulosic biomass can be completed in minutes, rather than weeks. The use of modern spectroscopy and chemometric techniques has shown promise as a rapid and cost effective alternative to traditional empirical techniques. This review serves as an overview of the compositional analysis techniques that have been developed for lignocellulosic biomass in an effort to highlight the motivation and migration towards rapid, accurate, and cost-effective data-driven chemometric methods. These rapid analysis techniques can potentially be used to optimize future biorefinery unit operations, where large quantities of lignocellulose are continually processed into products of high value.
A novel approach is presented for the synthesis of block-copolymers that resemble the architecture of a core cross-linked micelle. The polymers are synthesized from a combination of catalytic chain transfer polymerization (CCTP), thiol-Michael addition chemistry and reductive amination. A hydrophobic hyperbranched core is synthesized via CCTP of methyl methacrylate (MMA) and ethylene glycol dimethacrylate (EGDMA), which affords control over the polymer architecture and the degree of chain end-functionality. The vinyl unsaturations of the hyperbranched polymers are converted in nucleophilic pendant amines by thiol-Michael addition using cysteamine hydrochloride. A polysaccharide shell is grafted onto the hyperbranched core via reductive amination with dextran (DEX). The synthesized poly(MMA-co-EGDMA)-b-DEX polymers possess an amphiphilic character, are colloidally stable and resemble the topology of a core cross-linked micelle. The presented methodology provides a robust, modular, and tuneable approach towards the synthesis of amphiphilic core cross-linked micelle analogues.
Pneumonia remains the single leading cause of childhood death worldwide. Despite the commercial availability of multiple pneumococcal conjugate vaccines (PCVs), high dosage cost and supply shortages prevent PCV delivery to much of the developing world. The current work presents high-yield pneumococcal conjugates that are immunogenic in animals and suitable for use in human vaccine development. The 13-valent pneumococcal conjugate vaccine (PCV-13) investigated in this research incorporated serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F. Pneumococcal polysaccharides (PnPSs) and CRM197 carrier protein were produced and purified in-house, and used to prepare PnPS-CRM conjugates using unique, cyanide-free, in vacuo glycation conjugation methods. In vitro characterization confirmed the generation of higher molecular weight PnPS-CRM conjugates low in free protein. In vivo animal studies were performed to compare PnuVax's PCV-13 to the commercially available PCV-13, Prevnar®13 (Pfizer, USA). A boost dose was provided to all groups post-dose 1 at t = 14 days. Post-dose 2 results at t = 28 days showed that all 13 serotypes in PnuVax's PCV-13 were boostable. Per serotype IgG GMCs demonstrated that PnuVax's PCV-13 is immunogenic for all 13 serotypes, with 10 of the 13 serotypes statistically the same or higher than Prevnar®13 post-dose 2. As a result, the novel polysaccharide-protein conjugates developed in this work are highly promising for use in human PCV development. The in vacuo conjugation technique applied in this work could also be readily adapted to develop many other conjugate vaccines.
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