The recovery and purification of bioproducts is a key step that follows manufacture of proteins, peptides, and bioproducts. In the case of proteins and some types of bioproducts derived through fermentation or cell culture. Their recovery and purification (i.e., bioseparations) make-up more than 50% of manufacturing costs. An overlapping consideration is the impact of bio-physical properties and/or activity profiles on the recovery, fractionation, purification, and concentration of molecules and/or cells derived from fermentation or cell culture, and from catalytic or biocatalytic transformations.

Our laboratory combines sample analysis by liquid and gas chromatography, gavimetric biomass compositional analysis (NREL: https://www.nrel.gov/bioenergy/biomass-compositional-analysis.html), rheology, colligative solution properties gel electrophoresis, plating assays, gene sequencing, SEM, multiphoton laser confocal microscopy, rheology, and mass spectrometry to characterize molecules and properties of solutions derived from the experiments carried out in LORRE. The analytical procedures provide data required to validate first principles models and to generate databases for purposes of mechanistic analysis based of results and the literature by our research team, assisted by artificial intelligence (AI) and machine learning (ML) methods and software.

These capabilities are important for assessing the chemical make-up of plant lignocellulosic biomass and for determining material balances around biochemical and thermochemical processing of plant biomass. Transformation of agricultural bioproducts derived directly from soybean and corn is a major theme in our laboratory and is supported by the Indiana Soybean Alliance (ISA) and Indiana Corn Marketing Council (ICMC) (https://incornandsoy.org/). Current focus is on nutraceuticals from corn processing co-products, and sustainable aviation biofuel and heat transfer fluids from soybean. Analytical support for these areas utilizes the same instruments as outlined for analysis of biomolecules, but with different methods.

Sample cataloguing and preparation

Quantitative chemical analyses of liquid samples by HPLC and GC in addition to enzyme assays and gravimetric methods are central to being able to quantify biomass compositions, carbohydrates, organic acids, alcohols, and protein. Analytical capabilities represent expertise in compositional analysis and bio-physical property characterization of biomass and agricultural (lignocellulosic) materials, agricultural commodities and products, and rheological measurement of biomass slurries and highly concentrated proteins. Other analytical procures include liquid samples from fermentations (both aerobic and anaerobic), enzyme reactions, aqueous phase thermochemical reactions, anaerobic digesters, distillation condensates, and plant extracts.

In addition to standard analyses, we develop new HPLC analytical methods using ion exclusion, reverse phase, and ion exchange chromatography tailored to the identification and quantification of biomolecules of interest. Analytical capabilities are applied to bioseparations and membrane separations methods development applicable to process scale bioseparations for the manufacture of bioproduct and biopharmaceutical molecules and bioproducts.

HPLC analysis calibration

In addition to an array of wet-biological and chemical methods, BSL-2 laboratory facilities enable research on capture, concentration, and detection of food and other types of pathogens. Imaging instruments facilitate monitoring and validation of binding of labeled markers with microbial targets, proteins, and peptides and particulate structures of various micron, submicron, and molecular-scale sizes. In addition, our laboratory has developed specialized techniques for quantitating the movement of unlabeled proteins within matrices that provide simulated, in vitro subcutaneous, and intravitreal environments. Diffusion of injected therapeutic proteins are measured within these in vitro environments for development and validation of diffusional models. Doppler imaging (Physics laboratory of Professor David Nolte; (https://www.physics.purdue.edu/index.html) facilitates research on interactions between pathogens and immortalized mammalian cells and organoids grown in LORREs cell culture facility.

LORRE performs liquid-phase thermochemical reactions of biologically derived molecules (sugars, alcohols, etc.) over solid catalysts or in homogeneous reaction solutions at temperatures up to 260C. The HPLC and biomass analytical capabilities support measurement of product yields, rates, reaction selectivity, activation energy, and other engineering parameters necessary for reactor and catalyst design and scale-up.

These combined analytical capabilities provide an essential toolbox with which to carry out a range of research, and to develop new and scalable bioseparations. These include tangential flow filtration and liquid chromatographic methods. Overall, LORREs analytical capabilities and history in bioseparations development provide synergies that support overall research activities in our laboratory and with collaborators in other departments and disciplines at Purdue University.