We used continuous flow micro-devices as bioreactors for the production of a glycosylated pharmaceutical product (a monoclonal antibody). We cultured CHO cells on the surface of PMMA/PDMS micro-channels that had been textured by micromachining and coated with fibronectin. Three different micro-channel geometries (a wavy channel, a zigzag channel, and a series of donut-shape reservoirs) were tested in a continuous flow regime in the range of 3 to 6 μL min(-1). Both the geometry of the micro-device and the flow rate had a significant effect on cell adhesion, cell proliferation, and monoclonal antibody production. The most efficient configuration was a series of donut-shaped reservoirs, which yielded mAb concentrations of 7.2 mg L(-1) at residence times lower than one minute and steady-state productivities above 9 mg mL(-1) min(-1). These rates are at about 3 orders of magnitude higher than those observed in suspended-cell stirred tank fed-batch bioreactors.

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We report the design, fabrication, and characterization of novel, low-cost, and modular miniaturized nanofiber electrospinning sources for the scalable production of non-woven aligned nanofiber mats with low diameter variation. The devices are monolithic arrays of electrospinning emitters made via stereolithography; the emitters are arranged so each element has an independent line of sight to a rotating collector surface. Linear and zigzag emitter packing were evaluated using a PEO solution with the aim of maximizing the throughput of nanofibers with the smallest diameter and narrowest distribution. Current versus flowrate characterization of the devices showed that for a given flowrate a zigzag array produces more current per emitter than a linear array of the same emitter pitch and array size. In addition, the data demonstrate that larger and denser arrays have a net gain in flow rate per unit of active length. Visual inspection of the devices suggests uniform operation in devices with as many as 17 emitters with 300 μm inner diameter and 1.5 mm emitter gap. Well-aligned nanofiber mats were collected on a rotating drum and characterized; the 17-emitter device produced the same narrow nanofiber distribution (∼81 nm average diameter, ∼17 nm standard deviation) for all tested flow rates, which is strikingly different to the performance shown by 1-emitter sources where the average fiber diameter significantly increased and the statistics notably widened when the flowrate increases. Therefore, the data demonstrate that massively multiplexing the emitters is a viable approach to greatly increase the throughput of non-woven aligned nanofiber mats without sacrificing the statistics of the nanofibers generated. The production of dry nanofibers by the 17-emitter array is estimated at 33.0 mg min (1.38 mg min per mm of active length), which compares favorably with the reported multiplexed electrospinning arrays with emitters distributed along a line.

We report a proof-of-principle for the use of micro-devices as continuous bioreactors for the production of monoclonal antibodies. We culture CHO cells on the surface of PMMA "zigzag" channels textured with semi-spherical cavities coated with fibronectin, observing steady-state productivities 100 times higher than those observed in full scale systems.

The laser micro-spot welding process was studied to implement a sheet lamination process-based methodology for the fabrication of austenitic stainless steel scaffolds. AISI 302 sheets with a thickness of 254 μm were laser cut and laser welded. Experimental tests were carried out with different values of average laser power (i.e., 180, 200, and 220 W) and different exposure times (25, 50, 75, 100, 125 ms). The micro-spot welds were visually inspected according to the ISO 13919-1 Class B requirements. Spot welds were qualitatively characterized, and weld dimensions were measured (i.e., penetration depth, top, middle, and bottom width and the heat-affected zone (HAZ)) to identify the cross-sectional shape. Furthermore, process efficiencies (i.e., coupling, melting, and process) were studied. A seam welding model was adapted to calculate the required exposure time and was used to obtain a micro-spot weld to accomplish the quality requirements of the scaffold. A scaffold prototype was designed and manufactured using the selected parameters by experimental trials and using the mathematical model (i.e., a laser power of 220 W and an exposure time of 45 ms).