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When proteins in aqueous solutions are exposed to solid substrates, they adsorb due to the dynamic interplay of electrostatic, van der Waals, and hydration interactions and do so in a rather irreversible fashion, which makes protein recovery troublesome. Here, we use a gold electrode as the solid substrate and modulate the surface potential to systematically induce protein adsorption as well as partial desorption. We use different methods such as surface plasmon resonance, atomic force microscopy, and electrowetting and show that biasing the electrode to more negative potentials (by -0.4 V compared to the open-circuit potential at pH 6) results in an increased adsorption barrier of 6 kJ mol -1 for the negatively charged protein β-lactoglobulin. Further, we clearly demonstrate that this is due to an increased double layer potential of -0.06 V and an increase in hydration repulsion. This indicates that an electric potential can directly influence surface interactions and thus induce partial β-lactoglobulin desorption. These observations can be the basis for biosensors as well as separation technologies that use only one trigger to steer protein ad- and desorption, which is low in energy requirement and does not generate large waste streams, as is the case for standard protein separation technologies.

Microfluidic devices are an emerging technology for processing suspensions in e.g. medical applications, pharmaceutics and food. Compared to larger scales, particles will be more influenced by migration in microfluidic devices, and this may even be used to facilitate segregation and separation. In order to get most out of these completely new technologies, methods to experimentally measure (or compute) particle migration are needed to gain sufficient insights for rational design. However, the currently available methods only allow limited access to particle behaviour. In this review we compare experimental methods to investigate migration phenomena that can occur in microfluidic systems when operated with natural suspensions, having typical particle diameters of 0.1 to 10 μm. The methods are used to monitor concentration and velocity profiles of bidisperse and polydisperse suspensions, which are notoriously difficult to measure due to the small dimensions of channels and particles. Various methods have been proposed in literature: tomography, ultrasound, and optical analysis, and here we review and evaluate them on general dimensionless numbers related to process conditions and channel dimensions. Besides, eleven practical criteria chosen such that they can also be used for various applications, are used to evaluate the performance of the methods. We found that NMR and CSLM, although expensive, are the most promising techniques to investigate flowing suspensions in microfluidic devices, where one may be preferred over the other depending on the size, concentration and nature of the suspension, the dimensions of the channel, and the information that has to be obtained. The paper concludes with an outlook on future developments of measurement techniques., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Nowadays microfluidic devices are becoming popular for cell/DNA sorting and fractionation. One class of these devices, namely deterministic ratchets, seems most promising for continuous fractionation applications of suspensions (Kulrattanarak et al., 2008 [1]). Next to the two main types of particle behavior, zigzag and displacement motion as noted by the inventors (Huang et al., 2004 [2]) and (Inglis et al., 2006 [3]), we have shown recently the existence of a intermediate particle behavior, which we named 'mixed motion'. In this paper we formulate the hypothesis that the occurrence of mixed motion is correlated with anisotropy in the permeability of the obstacle array. This hypothesis we base on the comparison of experimental observations of mixed motion and the flow lane distribution as obtained from 2-D flow simulations., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Membrane processes are well-known for separating and fractionating suspensions in many industries, but suffer from particle accumulation on the membrane surface. Currently, there are new developments using microfluidic devices for cell/DNA sorting and fractionation. We anticipate these devices are also applicable to fractionation of polydisperse and concentrated suspensions (e.g. foods), and may potentially have fewer problems with particle accumulation compared to membranes. This review article presents an overview of relevant microfluidic devices. We focus on their performance with respect to concentrated suspensions, as one finds in food industry. We give quantitative estimates on their yield, selectivity, and the potential for large-scale application. From this evaluation follows that deterministic ratchets seem most promising.

Multiple phenomena are involved in conversions by immobilized biocatalysts. A paradox is identified between analytical desires on one hand and analytical boundary conditions on the other: while the study of interdependent phenomena would call for their simultaneous analysis in an integrated context, the available experimental options may impose a series of separate and dedicated analyses. From this analysis, bottlenecks in particle performance may be identified, if possible supported by a mechanistic model and performance criteria. Subsequently, a strategy for further biocatalyst development may be chosen. Finally, possibilities for future improvement of biocatalysts are discussed for various fields of research. Some examples of recent developments in enzyme and matrix characteristics, reactor operation, and micro-technology are discussed.

In a study of Assemblase, an industrial immobilized penicillin-G acylase, various electron microscopic techniques were used to relate intra-particle enzyme heterogeneity with the morphological heterogeneity of the support material at various levels of detail. Transmission electron microscopy was used for the study of intra-particle penicillin-G acylase distribution in Assemblase particles of various sizes; it revealed an abrupt increase in enzyme loading at the particle surface (1.4-fold) and in the areas (designated halo's) surrounding internal macro-voids (7.7-fold). Cryogenic field-emission scanning electron microscopy related these abrupt local enzyme heterogeneities to local heterogeneity of the support material by revealing the presence of dense top layers surrounding both the particle exterior and the internal macro-voids. Furthermore, it showed a very distinct morphological appearance of the halo. Most probably, all these regions contained relatively more chitosan than gelatin (the polymers Assemblase was constructed of), which suggested local polymer demixing during particle production. A basic thermodynamic line of reasoning suggested that a difference in hydrophilicity between the two polymers induced local demixing. In the future, thermodynamic knowledge on such polymer interactions resulting in matrix heterogeneity may be used as a tool for biocatalyst design.

Membrane emulsification is a promising and relatively new technique for producing emulsions. The purpose of this study was to better understand the influence of interfacial tension on droplet formation during membrane emulsification. Droplet formation experiments were carried out with a microengineered membrane; the droplet diameter and droplet formation time were studied as a function of the surfactant concentration in the continuous phase. These experiments confirm that the interfacial tension influences the process of droplet formation; higher surfactant concentrations lead to smaller droplets and shorter droplet formation times (until 10 ms). From drop volume tensiometer experiments we can predict the interfacial tension during droplet formation. However, the strong influence of the rate of flow of the to-be-dispersed phase on the droplet size cannot be explained by the predicted values. This large influence of the oil rate of flow is clarified by the hypothesis that snap-off is rather slow in the studied regime of very fast droplet formation.

In this study the influence of diffusion limitation on enzymatic kinetically controlled cephalexin synthesis from phenylglycine amide and 7-aminodeacetoxycephalosporinic acid (7-ADCA) was investigated systematically. It was found that if diffusion limitation occurred, both the synthesis/hydrolysis ratio (S/H ratio) and the yield decreased, resulting in lower product and higher by-product concentrations. The effect of pH, enzyme loading, and temperature was investigated, their influence on the course of the reaction was evaluated, and eventually diffusion limitation was minimised. It was found that at pH >or=7 the effect of diffusion limitation was eminent; the difference in S/H ratio and yield between free and immobilised enzyme was considerable. At lower pH, the influence of diffusion limitation was minimal. At low temperature, high yields and S/H ratios were found for all enzymes tested because the hydrolysis reactions were suppressed and the synthesis reaction was hardly influenced by temperature. The enzyme loading influenced the S/H ratio and yield, as expected for diffusion-limited particles. For Assemblase 3750 (the number refers to the degree of enzyme loading), it was proven that both cephalexin synthesis and hydrolysis were diffusion limited. For Assemblase 7500, which carries double the enzyme load of Assemblase 3750, these reactions were also proven to be diffusion limited, together with the binding-step of the substrate phenylglycine amide to the enzyme. For an actual process, the effects of diffusion limitation should preferably be minimised. This can be achieved at low temperature, low pH, and high substrate concentrations. An optimum in S/H ratio and yield was found at pH 7.5 and low temperature, where a relatively low reaction pH can be combined with a relatively high solubility of 7-ADCA. When comparing the different enzymes at these conditions, the free enzyme gave slightly better results than both immobilised biocatalysts, but the effect of diffusion limitation was minimal., (Copyright 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 80: 331-340, 2002.)