Biotechnological Manufacturing Research Articles

Lonza – hazardous flow chemistry for streamlined large scale synthesis

Lonza, a global leader in custom manufacturing, has been active for years in continuous fl ow operations using microreactor technologies, preferentially applied for hazardous reactions, and has core expertise in ozonolysis. Both key expertises have now been combined. Lonza is focused on the life sciences (bioresearch, pharma and biotech, nutrition, personal care, microbial control, agriculture and materials science). It has its headquarters in Basel, Switzerland with a total of 45 facilities and more than 11,000 employees. The product and service offering ranges from worldwide exclusive active substances to organic chemicals and intermediates manufactured in biotechnological and chemical processes. These substances are made for the chemical, pharmaceutical, agrochemical and food industries. Lonza is also strong in production, chemical and biotechnological research and development is another strength of Lonza. This includes profound know-how in organic syntheses and biotechnological processes, project management and the technology transfer from research and development into industrial-scale production. The main activities in this area are design, implementation and optimization of costcompetitive, safe and environmentally friendly chemical and biotechnological manufacturing processes for new fi ne chemicals and active pharmaceutical ingredients. One of Lonza’s core competencies is fi nding the optimal synthesis routes and processes for the industrial production of known compounds. As one of the recent innovative key technologies, Lonza re-designed chemical routes with the concept of fl ash chemistry in fl ow. Such highly intensifi ed fl ow processes most often need microreactors (Figures 1 and 2 ) because of their excellent heat transfer and mixing, exact control of residence time and segregation of feeds, together with small volume handling and robust design (pressure). Because of the high level of confi nement, the design is inherently safer. This paves the road to the intensifi ed mini-plant concept, for which the following drastic process intensifi cations were observed: lower reactor investment, less manpower, higher fl exibility, enhanced safety and faster change-over. This triggers the notion of new production concepts with the design of the “ factory of tomorrow ” . Currently, microreactors are restricted in use to the reaction part, to ensure high reaction yields and selectivity (Figure 3 ); in the future, this is foreseen as a fully integrated process including fl ow separations.

Best practices in heterotrophic high-cell-density microalgal processes: achievements, potential and possible limitations

Microalgae of numerous heterotrophic genera (obligate or facultative) exhibit considerable metabolic versatility and flexibility but are currently underexploited in the biotechnological manufacturing of known plant-derived compounds, novel high-value biomolecules or enriched biomass. Highly efficient production of microalgal biomass without the need for light is now feasible in inexpensive, well-defined mineral medium, typically supplemented with glucose. Cell densities of more than 100 g l−1 cell dry weight have been achieved with Chlorella, Crypthecodinium and Galdieria species while controlling the addition of organic sources of carbon and energy in fedbatch mode. The ability of microalgae to adapt their metabolism to varying culture conditions provides opportunities to modify, control and thereby maximise the formation of targeted compounds with non-recombinant microalgae. This review outlines the critical aspects of cultivation technology and current best practices in the heterotrophic high-cell-density cultivation of microalgae. The primary topics include (1) the characteristics of microalgae that make them suitable for heterotrophic cultivation, (2) the appropriate chemical composition of mineral growth media, (3) the different strategies for fedbatch cultivations and (4) the principles behind the customisation of biomass composition. The review confirms that, although fundamental knowledge is now available, the development of efficient, economically feasible large-scale bioprocesses remains an obstacle to the commercialisation of this promising technology.

BIOPREPARATIONS IN BIOORGANIC AGRICULTURE

The results of investigations of Ukrainian and Russian scientists- microbiologists as for creation of microbic preparations and their ap¬plication in bioorganic agriculture are generalized. The major factors and the scheme of biotechnological manufacturing of specified biologi¬cal products are considered. It has been shown, that one of the most important properties of the preparation is the form of manufacturing. Searching of universal fills is very actual as well as different updatings of biological products which provide maximal adaptibility of biological agents to specific conditions, depending on regions. For the first time gel, lignin and vermiculturas forms of biological products based on new strains of nitrogen fixing bacteria have been developed by Ukrainian scientists.

Probe-Independent and Direct Quantification of Insulin mRNA and Growth Hormone mRNA in Enriched Cell Preparations

Task division in multicellular organisms ensures that differentiated cell types produce cell-specific proteins that fulfill tasks for the whole organism. In some cases, the encoded mRNA species is so abundant that it represents a sizeable fraction of total mRNA in the cell. In this study, we have used a probe- and primer-free technique to quantify such abundant mRNA species in order to assess regulatory effects of in vitro and in vivo conditions. As a first example, we were able to quantify the regulation of proinsulin mRNA abundance in beta-cells by food intake or by the glucose concentration in tissue culture. The second example of application of this technique is the effect of corticosteroids on growth hormone mRNA in enriched somatrotrophs. It is anticipated that other examples exist in which measurement of very abundant mRNAs in dedicated cells will help to understand biological processes, monitor disease states, or assist biotechnological manufacturing procedures.

Size Shrinkage of Methacrylate-based Terpolymer Latexes Synthesized by Free Radical Polymerization: Kinetics and Influence of Main Reaction Parameters

Monodisperse PMMA-based terpolymer particles were synthesized by surfactant-free free radical emulsion polymerization. Particles with a wide and controllable range of size and polymer content were prepared by varying monomer amount, reaction temperature and initiator concentration. A kinetic study of the evolution of the terpolymer colloid composition and morphology reveals a mechanism of fast homogeneous nucleation. Copolymerization of the three monomers yields particles whose composition changes continuously during the reaction, which impacts polymer properties. Temperature and monomer amount are key parameters, particularly in the nucleation and growth stages of the reaction. A linear relationship between the particle size and reaction temperature is observed. This is attributed to the fact that the concentration of small, nucleated primary particles with high surface area, initiated at higher temperatures, is too large for efficient stabilization and hence they are more prone to aggregation. The linear relationship between the volume of the colloids and the ratio of main monomer to water in the reaction batch is due to the growth of the particles that continues while there is MMA available. The initiator concentration is not significant in terms of size and concentration of methacrylate-based colloids since primary particles are formed very early during polymerization and they are not dependent on the number of growing chains. The resulting “tailor-made” latexes are promising for a number of unique biotechnological and IC manufacturing applications.

Modeling flocculation of biological cells

Biotechnological manufacturing processes often include a bioreactor harvesting step which involves separation of cells, cell debris and other particulate matter from the suspending medium which frequently contains the product of interest. Tangential flow microfiltration is used to separate the particulate matter from the suspending medium. The efficiency of this microfiltration step is often improved by increasing the particle size through flocculation prior to microfiltration. Here flocculation of yeast suspensions using a cationic polyelectrolyte has been investigated. A self-similar steady state floc size distribution is obtained experimentally. The experimental results may be modeled using a population balance approach. The dimensionless floc size distribution is found to be independent of the particulate volume fraction, initial particle size and shear rate during flocculation for the ranges of these variables considered. However, the actual floc size distribution does depend upon these parameters. In addition, the approach to the self-similar steady state floc size distribution depends upon the particulate volume fraction. At higher particulate volume fractions, the volume average floc size is found to increase during flocculation, pass through a maximum and then decrease to the final steady state value. However, at lower particle volume fractions, a maximum in the volume average floc size is not observed. Flocculation of cells leads to an increase in the volume average particle diameter. However, it also leads to a broadening of the particle size distribution. Consequently, when determining the improvement in permeate flux for tangential flow microfiltration, it is essential to include the effects of an increased average particle diameter and a broadened particle size distribution.

The Changing Landscape of Regulatory Control of Biological Medicines

This paper focuses on the rapid growth in biological medicines in recent years brought about by advances in molecular biology and biotechnological manufacturing techniques. It looks at the implications for regulatory bodies and research institutions concerned with quality control of biologicals; and discusses ways in which the industry/regulatory/research partnership is evolving in response to the new situation. It argues that, although change is being managed relatively successfully in some areas of regulatory control (devolution to industry, use of new technologies, international harmonization), special measures may be needed in some other areas. These include: needs of the developing world for biologicals quality control; sustaining long-term research on safety issues and test methodology; major social and ethical issues raised by new treatment and diagnostic opportunities.

Experimental approaches to guarantee minimal risk of potential virus in purified monoclonal antibodies.

Regarding biological products, increasing awareness of potential side effects have placed great importance not only at protein purity regarding other proteins but on the removal of biologicals such as DNA and especially virus the importance of which may not be known. Monoclonal antibodies (Mab) have come to be an important class of molecules obtained from hybridoma cells, i.e., nonrecombinant cells in culture. It has been noted during the last years, that with rare exceptions hybridoma cell lines contain retrovirus like particles. The infectious nature of the EM-visible particles has been tested for, however, in most cases not been substantiated. In order to bring these valuable biological reagents, Mab's, to good use in man for imaging or therapy, the remaining concern about a potential retroviral infection has to be reduced to an acceptable minimum. We describe experimental approaches for the validation of chromatographic and ultrafiltration steps used in the production of monoclonal antibodies to remove and inactivate murine retrovirus. Present day biotechnological manufacturing processes have been devised incorporating a number of strategic preventive measures that have found wide spread acceptance. They permit to answer the question: how can a potentially harmful infection by an unknown virus be excluded. Knowledge of the efficacy of purification steps to clear infectious model virus is fundamental to devise biotechnological manufacturing processes yielding a purified antibody for use in man.