Cross‐flow filtration of fine suspensions through microsieves occurs in microprocessing. The interaction of particles with surfaces in microenvironments has been extensively studied, but predominantly in monolayers and not with an eye to microfiltration. Here, we introduce a microfiltration model that pertains to particles that might be seen as fine in a macroscopic environment, but are large enough to intrude significantly into the shear layer of a microchannel. Thus, particle accumulation upon the sieve couples the steady‐state filtrate flux and the suspension flow through the microchannel that feeds the sieve. We envision and create a stable, stationary multilayer of particles whose thickness is shear‐limited and we identify and verify the structure and parameters that limit steady filtration in this environment. At first, a packed bed of particles forms, growing into and regulated by the micro channel's shear flow. A critical shear stress is shown to determine the thickness of the bed, seen as a stationary and stable multilayer of particles through which filtration may occur. As the bed thickens, at the expense of channel area for suspension flow, surface shear stress increases until no further particle adherence is possible. We built a simple example using hard noninteracting polymer microspheres and conducted cross‐flow filtration experiments over Aquamarijn™ microsieves (uniform pore size of 0.8 μm). We observed a steady cake‐layer thickness and because of the simple geometry afforded by uniform spheres, we could approximate the force balance, cake resistance, and filtration rate from first principles. The good fit of our data to the proposed mechanism lays a firm basis for the semiquantitative analysis of the behavior of more complex suspensions. © 2018 American Institute of Chemical Engineers AIChE J, 65: 207–213, 2019

Steady state crossflow microfiltration (CMF) is an important and often necessary means of particle separation and concentration for both industrial and biomedical processes. The factors controlling the performance of CMF have been extensively reviewed. A major factor is transmembrane pressure (TMP). Because microchannels have small height, they tend to have high pressure gradients in the feed-flow direction. In the extreme, these gradients may even reverse the pressure across the membrane (inciting backflow). It is therefore desirable to compensate for the effect of feed-flow on the TMP, aiming at constant transmembrane pressure (cTMP) at a value which maximizes filtrate flux. This is especially critical during filtration of deformable particles (e.g. erythrocytes) through low intrinsic resistance membranes. Filtration flux is generally taken to be directly proportional to TMP, with pressure drop along the channel decreasing in the flow direction. A co-current flow of filtrate in a suitably designed filtrate collecting channel is shown to allow the TMP to remain constant and permit the sieving surface to perform optimally, permitting up to twice as much filtration over that of a naïve configuration. Manipulation of the filtrate channel may be even more beneficial if it prevents backflow that might otherwise occur at the end of a sufficiently long channel. Experiments with erythrocyte suspensions, reported here, validate these concepts.

Crossflow microfiltration of plasma from blood through microsieves in a microchannel is potentially useful in many biomedical applications, including clinically as a wearable water removal device under development by the authors. We report experiments that correlate filtration rates, transmembrane pressures (TMP) and shear rates during filtration through a microscopically high channel bounded by a low intrinsic resistance photolithographically-produced porous semiconductor membrane. These experiments allowed observation of erythrocyte behavior at the filtering surface and showed how their unique deformability properties dominated filtration resistance. At low filtration rates (corresponding to low TMP), they rolled along the filter surface, but at higher filtration rates (corresponding to higher TMP), they anchored themselves to the filter membrane, forming a self-assembled, incomplete monolayer. The incompleteness of the layer was an essential feature of the monolayer’s ability to support sustainable filtration. Maximum steady-state filtration flux was a function of wall shear rate, as predicted by conventional crossflow filtration theory, but, contrary to theories based on convective diffusion, showed weak dependence of filtration on erythrocyte concentration. Post-filtration scanning electron micrographs revealed significant capture and deformation of erythrocytes in all filter pores in the range 0.25 to 2 μm diameter. We report filtration rates through these filters and describe a largely unrecognized mechanism that allows stable filtration in the presence of substantial cell layers.

Peristaltic pumps rely on constant compression of elastomeric tubing from which particles may be shed, a phenomenon known as spallation. We studied spallated particles on microfluidic filtration devices with photolithographically prepared micron-level pore fields. Filtration of ultra-pure water through these pores was analyzed using either the usual peristaltic pump or a reciprocating pair of syringe pumps. Using syringe pumps, transmembrane pressure (TMP) values during filtration at 2.5 cm3/min revealed steady filtration for over 80 minutes at 2.3 mmHg. Using the peristaltic pump, TMP was never stable, increasing to approximately 11 mmHg during the first 10 minutes. Pore plugging was the culprit, evidenced by post-perfusion microphotography.

Knowledge of dynamics of shift of fluid volume between intra- and extravascular compartments during hemodialysis (HD) is important for managing HD treatment to help patients approach dry weight without hypotension. The Relative blood volume (RBV) monitor indicates change in plasma volume based on the difference between ultrafiltration rate (UFR) and plasma refilling rate (PRR) during HD. However, the absolute value of PRR cannot be obtained from RBV. The aim of this study was to investigate whether fluid transport from the interstitial to blood spaces can be quantitatively analyzed with a two compartments model. 14 patients (30 measurements) were studied. RBV using a blood volume monitor (BVM, Fresenius) and calf extracellular volumes (ECV) by calf bioimpedance device (Hydra 4200, Xitron) were continuously measured during HD. A mathematic model was established with unknown transport coefficients (k1, k2, α, β, γ, δ) and these coefficients were estimated using a Least Squares Optimization algorithm by fitting from experimental data. A high correlation (R2>0.8) between experimental data and calculation by the model were observed in both RBV and ECV measurements. Coefficients k1 and δ significantly differed with different degree of hydration. This model provides parameters which can used to understand relationships between degree of hydration and refilling rate.

Co-amplification of transgenes using the dihydrofolate reductase/methotrexate (DHFR/MTX) system is a widely used method for the isolation of Chinese hamster ovary (CHO) cell lines that secrete high levels of recombinant proteins. A bottleneck in this process is the stepwise selection for MTX resistant populations; which can be slow, tedious and erratic. We sought to speed up and regularize this process by isolating dhfr(-) CHO cell lines capable of integrating a transgene of interest into a defined chromosomal location that supports a high rate of gene amplification. We isolated 100 independent transfectants carrying a gene for human adenosine deaminase (ada) linked to a phi C31 attP site and a portion of the dihydrofolate reductase (dhfr) gene. Measurement of the ada amplification rate in each transfectant using Luria-Delbruck fluctuation analysis revealed a wide clonal variation; sub-cloning showed these rates to be heritable. Site directed recombination was used to insert a transgene carrying a reporter gene for secreted embryonic alkaline phosphatase (SEAP) as well as the remainder of the dhfr gene into the attP site at this location in several of these clones. Subsequent selection for gene amplification of the reconstructed dhfr gene in a high ada amplification candidate clone (DG44-HA-4) yielded reproducible rates of seap gene amplification and concomitant increased levels of SEAP secretion. In contrast, random integrations of the dhfr gene into clone HA-4 did not yield these high levels of amplification. This cell line as well as this method of screening for high amplification rates may prove helpful for the reliable amplification of recombinant genes for therapeutically or diagnostically useful proteins. (C) 2013 Elsevier B.V. All rights reserved.

s are only available online, free of charge, under www.karger.com/doi/10.1159/000345376 30th Annual Meeting of the International Society of Blood Purification (ISBP) September 6–8, 2012, Yokohama, Japan

Background: Although prior studies have shown that frequent hemodialysis (HD) can lead to improved control of dry weight in end-stage renal disease patients, there are no clinical studies examining whether this can improve blood pressure (BP) control and can also shorten the dialysis time needed to achieve satisfactory removal of small molecules. Several models of wearable dialysis systems are now under various stages of development. These devices present the possibility of hemodialyzing patients to their dry weights. We have built a prototype of a wearable ultrafiltration (UF) device that can provide daily UF. Apart from better fluid control, we hypothesize that separating HD from UF will result in better BP control, and adequate weekly small molecule removal could be achieved with a decreased duration of dialysis. We tested the hypothesis in current HD patients using conventional dialysis equipment. Methods: Thirteen patients were selected from a large urban HD center. The experimental period consisted of 4 weeks of daily UF (4 days/week of UF alone and 2 days/week of HD with UF). The duration of the HD sessions was increased by 15–30 min to maintain weekly standard Kt/V >2.0. The patients were then returned to their conventional 3 days/week of HD with UF and studied for 4 weeks. Predialysis BPs, interdialytic weight gains, and Kt/V results of the experimental and return periods were compared with those of the 3-month control period. No changes were made in antihypertensive or other medication during the study. Results: During the experimental period, mean arterial pressure decreased from 110 to 95 mm Hg (p < 0.001), systolic BP from 158 to 136 mm Hg (p < 0.001), while interdialytic weight gains were reduced from 3.25 to 1.21 liters (p < 0.0001). During the experimental period, weekly standard Kt/V of 2.16 was achieved in 8.24 h/week of HD, as compared to 11.14 h/week. Conclusions: Volume control with daily UF results in improved BP control and, by separating the UF function from HD, adequate weekly standard Kt/V >2 can be achieved with twice weekly HD.

We have designed a novel, low energy platelet-rich-plasma (PRP) separator capable of producing 50 mL of PRP in 30 min, intended for military and emergency applications. Blood flows over a 3 mm length of sieve at high rates of shear. A plasma-platelet filtrate passes through the sieve’s pores while erythrocytes remain. The filtrate is flowed over a second 3 mm length of smaller-pored sieve that withdraws plasma. Bulk blood volume is maintained by returning platelet-free plasma to the erythrocyte pool, enabling a nearly complete multi-pass platelet extraction. The total percentage of platelets extracted is: \( \theta (T) = 1 - exp\left( {\frac{{ - {V_f}(T){\phi_P}}}{V}} \right) \) where V is the original plasma volume, V f (T) is the total filtered volume, and ϕ P is platelet passage ratio (filtrate concentration/bulk average concentration) taken to be constant. Maximum θ(T) occurs at maximum V f (T) × ϕ P Test microsieves, 3 mm long × 3 mm wide, were used. ϕ P values measured at various filtrate flow rates (20–100uL/min) and utilizing various filter pore sizes (1.2–3.5 μm), was as high as 150 %. Maximum V f (T) × ϕ P was achieved utilizing the 3.5um filters at the highest flow rate, 100 uL/min. Erythrocyte leakages were always below 2,000/uL, far below the allowable limit stipulated by the American Association of Blood Banking. These data imply that a 13.7 cm2 filter area is sufficient to achieve the target separation of 50 mL of platelet concentrate in 30 min. The filtration cartridge would consist of multiple microporous strips of 3 mm width arranged in parallel so that each element would see the conditions used in the prototype experiments presented here. Other microfiltration schemes suggest no method of scaling to practical levels.

253 30th Annual Meeting of the International Society of Blood Purification (ISBP) September 6–8, 2012, Yokohama, Japan Guest Editors: Tadao Akizawa (Tokio); Robert Zietse (Rotterdam) (available online only) 353 Erratum 364 Acknowledgement to the Reviewers

Wearable blood processing devices offer an attractive solution to problems inherent in clinic-based, intermittent end-stage renal disease therapies. What is involved in transitioning even a part of the current clinic-based population to ambulatory therapy has not been clearly enumerated. This paper addresses what a first-generation wearable device might accomplish, how issues of safety will need to be addressed, and what will make the device attractive to, and manageable by, the patient. Medical, technological, and economic issues are identified.

Many points of reference have been used to compare and rationalize extracorporeal end-stage renal disease therapy. We address a specific part of the subject: the effect of the delivery schedule on a predetermined dose of dialysis, e.g., weekly Kt/V. Steady (time-invariant) application of dialysis absolutely minimizes time-averaged and peak concentrations of any extractable solute. However, such dosing is often impractical; we assess the effectiveness of achievable slow regimens relative to steady dosing, using the single-pool approximation, applicable to slow regimens. Dose scheduling has been previously considered. We combine and discuss prior observations and establish continuous dosing as an easily quantifiable reference point, and we emphasize fundamental patterns common to different schedules. Thus, we enable rapid comparison of the many "slow" dialysis regimens presently under consideration using two intuitive parameters to encompass dialysis dosing: intermittency and intensity. These parameters define any repetitive dialysis pattern. A method for evaluating any combination of them is given with formulae, graphically, and with examples. Intermittency increases average solute concentration only slightly, but the frequency and spacing of intermittent treatments strongly affect peak solute concentrations. With steadier solute removal, cycling of solvent (water) stores is likely to remain the dominant source of disequilibrium in patients.