Using Passive Microrheology to Measure the Evolution of the Rheological Properties of NIST mAb Formulations during Adsorption to the Air-Water Interface.

The development of novel protein-based therapeutics, such as monoclonal antibodies (mAbs), is often limited due to challenges associated with maintaining the stability of these formulations during manufacturing, storage, and clinical administration. An undesirable consequence of the instability of protein therapeutics is the formation of protein particles. MAbs can adsorb to interfaces and have the potential to undergo partial unfolding as well as to form viscoelastic gels. Further, the viscoelastic properties may be correlated with their aggregation potential. In this work, a passive microrheology technique was used to correlate the evolution of surface adsorption with the evolution of surface rheology of the National Institute of Standards and Technology (NIST) mAb reference material (NIST mAb) and interface-induced subvisible protein particle formation. The evolution of the surface adsorption and interfacial shear rheological properties of the NIST mAb was recorded in four formulation conditions: two different buffers (histidine vs phosphate-buffered saline) and two different pHs (6.0 and 7.6). Our results together demonstrate the existence of multiple stages for both surface adsorption and surface rheology, characterized by an induction period that appears to be purely viscous, followed by a sharp increase in protein molecules at the interface when the film rheology is viscoelastic and ultimately a slowdown in the surface adsorption that corresponds to the formation of solid-like or glassy films at the interface. When the transitions between the different stages occurred, they were dependent on the buffer/pH of the formulations. The onset of these transitions can also be correlated to the number of protein particles formed at the interface. Finally, the addition of polysorbate 80, an FDA-approved surfactant used to mitigate protein particle formation, led to the interface being surfactant-dominated, and the resulting interface remained purely viscous.

Ethanol organosolv pretreatment of sugarcane bagasse assisted by organic acids and supercritical carbon dioxide.

The scCO2-assisted organosolv pretreatment of sugarcane bagasse was carried out using aqueous ethanol and organic acid catalysts. Variables involved were temperature (150-190 °C), time (0-60 min), type of catalyst (acetic, citric, and oxalic acids), amount of CO2 (25-50 g), ethanol titer in water (0-80 vol%), and catalyst concentration (0.5 to 1.5 % w·v-1). The best delignification (86 wt%) and glucan retention (89 wt%) were achieved at 170 °C for 15 min using 60 vol% ethanol in water, 1 wt% oxalic acid, and 25 g CO2. Organic acid esterification was a limitation for pretreatment operations using ethanol titers above 60 vol%. Enzymatic hydrolysis of pretreated materials at 1 % (w·v-1) glucans released 74.3 ± 0.2 % glucose in 96 h using Cellic CTec3 (Novozymes) at 9.89 FPU·gglucans-1. The concentrated pretreatment liquor allowed lignin recovery by water precipitation in high yields, while the aqueous supernatant contained low levels of fermentation inhibitors.

Supercritical Fluids: A Promising Technique for Biomass Pretreatment and Fractionation.

Lignocellulosic biomasses are primarily composed of cellulose, hemicelluloses and lignin and these biopolymers are bonded together in a heterogeneous matrix that is highly recalcitrant to chemical or biological conversion processes. Thus, an efficient pretreatment technique must be selected and applied to this type of biomass in order to facilitate its utilization in biorefineries. Classical pretreatment methods tend to operate under severe conditions, leading to sugar losses by dehydration and to the release of inhibitory compounds such as furfural (2-furaldehyde), 5-hydroxy-2-methylfurfural (5-HMF), and organic acids. By contrast, supercritical fluids can pretreat lignocellulosic materials under relatively mild pretreatment conditions, resulting in high sugar yields, low production of fermentation inhibitors and high susceptibilities to enzymatic hydrolysis while reducing the consumption of chemicals, including solvents, reagents, and catalysts. This work presents a review of biomass pretreatment technologies, aiming to deliver a state-of-art compilation of methods and results with emphasis on supercritical processes.