In situ NIR spectroscopy monitoring of plasmid production processes effect of producing strain, medium composition and the cultivation strategy

BACKGROUNDWhile the pharmaceutical industry keeps an eye on plasmid DNA production for new generation gene therapies, real-time monitoring techniques for plasmid bioproduction are as yet unavailable. This work shows the possibility of in situ monitoring of plasmid production in Escherichia coli cultures using a near infrared (NIR) fiber optic probe. RESULTSPartial least squares (PLS) regression models based on the NIR spectra were developed for predicting bioprocess critical variables such as the concentrations of biomass, plasmid, carbon sources (glucose and glycerol) and acetate. In order to achieve robust models able to predict the performance of plasmid production processes, independently of the composition of the cultivation medium, cultivation strategy (batch versus fed-batch) and E. coli strain used, three strategies were adopted, using: (i) E. coliDH5 cultures conducted under different media compositions and culture strategies (batch and fed-batch); (ii) engineered E. coli strains, MG1655endArecApgi and MG1655endArecA, grown on the same medium and culture strategy; (iii) diverse E. coli strains, over batch and fed-batch cultivations and using different media compositions. PLS models showed high accuracy for predicting all variables in the three groups of cultures. CONCLUSIONNIR spectroscopy combined with PLS modeling provides a fast, inexpensive and contamination-free technique to accurately monitoring plasmid bioprocesses in real time, independently of the medium composition, cultivation strategy and the E. coli strain used.

From weak Allee effect to no Allee effect in Richards’ growth models

Population dynamics have been attracting interest since many years. Among the considered models, the Richards’ equations remain one of the most popular to describe biological growth processes. On the other hand, Allee effect is currently a major focus of ecological research, which occurs when positive density dependence dominates at low densities. In this chapter, we propose the dynamical study of classes of functions based on Richards’ models describing the existence or not of Allee effect. We investigate bifurcation structures in generalized Richards’ functions and we look for the conditions in the (β, r) parameter plane for the existence of a weak Allee effect region. We show that the existence of this region is related with the existence of a dovetail structure. When the Allee limit varies, the weak Allee effect region disappears when the dovetail structure also disappears. Consequently, we deduce the transition from the weak Allee effect to no Allee effect to this family of functions. To support our analysis, we present fold and flip bifurcation curves and numerical simulations of several bifurcation diagrams.

Explosion birth and extinction: Double big bang bifurcations and Allee effect in Tsoularis-Wallace's growth models

This work concerns dynamics and bifurcations properties of a new class of continuous-defined one-dimensional maps: Tsoularis-Wallace's functions. This family of functions naturally incorporates a major focus of ecological research: the Allee effect. We provide a necessary condition for the occurrence of this phenomenon of extinction. To establish this result we introduce the notions of Allee's functions, Allee's effect region and Allee's bifurcation curve. Another central point of our investigation is the study of bifurcation structures for this class of functions, in a three-dimensional parameter space. We verified that under some sufficient conditions, Tsoularis-Wallace's functions have particular bifurcation structures: the big bang and the double big bang bifurcations of the so-called "box-within-a-box" type. The double big bang bifurcations are related to the existence of flip codimension-2 points. Moreover, it is verified that these bifurcation cascades converge to different big bang bifurcation curves, where for the corresponding parameter values are associated distinct kinds of boxes. This work contributes to clarify the big bang bifurcation analysis for continuous maps and understand their relationship with explosion birth and extinction phenomena.