Recovery Of Biological Products Research Articles

Development, characterization and application of 3D printed adsorbents for in situ recovery of taxadiene from microbial cultivations

Microbial cell factories are an attractive alternative to produce high-value natural products using sustainable processes. However, product recovery is one of the main challenges to reduce production cost and make these technologies economically interesting. In this work, new resins were formulated to 3D print hydrophobic adsorbents for the recovery of biologics from microbial cultivations. Benzyl methacrylate (BEMA) and butyl methacrylate (BUMA) were selected as functional monomers suitable for the adsorption of hydrophobic compounds. Pore morphology was tailored through the inclusion of pore forming agents (porogens) in the resin. Different porogens and porogen concentrations were evaluated resulting in materials with different porous networks. Sudan 1 and the anticancer drug paclitaxel were employed as model compounds to test the adsorption performance of hydrophobic and terpene molecules onto the developed 3D printed materials. The material with greatest adsorption capacity was obtained using BEMA monomer with 40 % (v/v) porogen (BEMA40). The performance of BEMA40 to recover taxadiene from small-scale (5 mL) Saccharomyces cerevisiae cultivations was tested and compared with commercial Diaion HP-20 beads. Taxadiene titres on BEMA40 (46 ± 2 mg/L) and Diaion HP-20 (54 ± 4 mg/L) were comparable, with no taxadiene detected in the cells and cell-free media, suggesting near 100 % taxadiene partition on the adsorbents. Compared to commercial beads, 3D printed adsorbents can be customized with adjustments in the resin formulation, are well adaptable to diverse bioreactor types, do not clog sampling ports and columns and are easier to handle during post processing. The results of this work demonstrate the potential of 3D printing to fabricate hydrophobic interaction adsorbent materials and their application in the recovery of biological products.

Insulin purification-Innovation continuum via synthesis of fundamentals, technology, and modeling.

Advancement in all disciplines (art, science, education, and engineering) requires a careful balance of disruption and advancement of classical techniques. Often technologies are created with a limited understanding of fundamental principles and are prematurely abandoned. Over time, knowledge improves, new opportunities are identified, and technology is reassessed in a different light leading to a renaissance. Recovery of biological products is currently experiencing such a renaissance. Crystallization is one example of an elegant and ancient technology that has been applied in many fields and was employed to purify insulins from naturally occurring sources. Crystallization can also be utilized to determine protein structures. However, a multitude of parameters can impact protein crystallization and the "hit rate" for identifying protein crystals is relatively low, so much so that the development of a crystallization process is often viewed as a combination of art and science even today. Supplying the worldwide requirement for insulin (and associated variants) requires significant advances in process intensification to support scale of production and to minimize the overall cost to enable broader access. Expanding beyond insulin, the increasing complexity and diversity of biologics agents challenge the current purification methodologies. To harness the full potential of biologics, there is a need to fully explore a broader range of purification technologies, including nonchromatographic approaches. This impetus requires one to challenge and revisit the classical techniques including crystallization, chromatography, and filtration from a different vantage point and with a new set of tools, including molecular modeling. Fortunately, computational biophysics tools now exist to provide insights into mechanisms of protein/ligand interactions and molecular assembly processes (including crystallization) that can be used to support de novo process development. For example, specific regions or motifs of insulins and ligands can be identified and used as targets to support crystallization or purification development. Although the modeling tools have been developed and validated for insulin systems, the same tools can be applied to more complex modalities and to other areas including formulation, where the issue of aggregation and concentration-dependent oligomerization could be mechanistically modeled. This paper will illustrate a case study juxtaposing historical approaches to insulin downstream processes to a recent production process highlighting the application and evolution of technologies. Insulin production from Escherichia coli via inclusion bodies is an elegant example since it incorporates virtually all the unit operations associated with protein production-recovery of cells, lysis, solubilization, refolding, purification, and crystallization. The case study will include an example of an innovative application of existing membrane technology to combine three-unit operations into one, significantly reducing solids handling and buffer consumption. Ironically, a new separations technology was developed over the course of the case study that could further simplify and intensify the downstream process, emphasizing and highlighting the ever-accelerating pace of innovation in downstream processing. Molecular biophysics modeling was also employed to enhance the mechanistic understanding of the crystallization and purification processes.

Microfluidic techniques help expand applications for all-aqueous emulsion droplets

Advances in forming and stabilizing droplets are poised to allow for further development in applications such as purification, cell encapsulation, partitioning, and recovery of biological products.

Process Intensification for Production and Recovery of Biological Products

Bioprocesses are important biological reactions which need several sophisticated methods and equipment to produce many novel and important compounds which some of them are traditionally produced by synthetic chemical reactions. In bioprocesses, the products are often produced in a dilute environment and finally they require a high purity. Because of that, downstream processes are usually included a large number of separation steps. Size and capital costs of the equipment are two main limitations of using bioprocesses at industrial scale. Bioprocess intensification by minimizing, substitution, moderation and simplification of the methods and equipment, drastically leads to sustainable processes. This study looks at intensification of the emerging equipment and operational methods and their advantages to lead smaller and cleaner bioprocess plants which in turn, increases production efficiency and quality and decreases byproducts formation, capital cost and energy consumption.

Recovery of Biological Products in Aqueous Two Phase Systems

Recovery of Biological Products in Aqueous Two Phase Systems

The Effect of Additives and Impurities on the Partition of Ethanol into n‐Decanol from Aqueous Solutions

AbstractThe recovery of biological products from fermentation media by direct liquid‐liquid extraction is complicated by the presence of impurities. These may comprise either side products of the bio‐transformation or residual species from the pre‐fermentation substrates. As a corollary to the detrimental effect of impurities upon extraction, addition of reagents, for example salts, can be used to enhance the extraction and mitigate to some extent against these. This paper describes an experimental study of the partitioning of ethanol from aqueous solutions into 1–decanol. Specific groups of impurities and additives were identified, comprsing: electrolyte compounds (including acids, bases, and salts), ionic and non‐ionic surfactants, commercial de‐emulsifiers, organic acids, polysaccharide compounds (i.e. sugars and starch), protein, biomass (including yeast and bacteria), and 1–propanol. A number of thermodynamic models for predicting the effect of each additive on the phase distribution behaviour of ethanol were screened and tested. The additives considered in this paper are confined to those of relatively simple chemical structure and included 1–propanol, oleic acid, glucose, sodium chloride, sodium hydroxide, calcium chloride, ammonium chloride, ammonium sulphate, and hydrochloric acid. These were added in controlled amounts to the principal extraction system components and the effect upon the partition coefficients of ethanol determined.