Search for: All records

Abstract We report the integration of 3D printing, electrobiofabrication, and protein engineering to create a device that enables near real‐time analysis of monoclonal antibody (mAb) titer and quality. 3D printing was used to create the macroscale architecture that can control fluidic contact of a sample with multiple electrodes for replicate measurements. An analysis “chip” was configured as a “snap‐in” module for connecting to a 3D printed housing containing fluidic and electronic communication systems. Electrobiofabrication was used to functionalize each electrode by the assembly of a hydrogel interface containing biomolecular recognition and capture proteins. Specifically, an electrochemical thiol oxidation is used to assemble a thiolated polyethylene glycol hydrogel, that in turn is covalently coupled to either a cysteine‐tagged protein G that binds the antibody's Fc region or a lectin that binds the glycans of target mAb analytes. We first show the design, assembly, and testing of the hardware device. Then, we show the transition of a step‐by‐step sensing methodology (e.g., mix, incubate, wash, mix, incubate, wash, measure) into the current method where functionalization, antibody capture, and assessment are performed in situ and in parallel channels. Both titer and glycan analyses were found to be linear with antibody concentration (to 0.2 mg/L). We further found the interfaces could be reused with remarkably similar results. Because the interface assembly and use are simple, rapid, and robust, we suggest this assessment methodology will be widely applicable, including for other biomolecular process development and manufacturing environments. 

A library of soft, stretchable, strain-insensitive bioelectronics was made using brittle interfacial materials. 

Abstract Redox provides unique opportunities for interconverting molecular/biological information into electronic signals. Here, the fabrication of a 3D‐printed multiwell device that can be interfaced into existing laboratory instruments (e.g., well‐plate readers and microscopes) to enable advanced redox‐based spectral and electrochemical capabilities is reported. In the first application, mediated probing is used as a soft sensing method for biomanufacturing: it is shown that electrochemical signal metrics can discern intact mAbs from partially reduced mAb variants (fragmentation), and that these near‐real‐time electrical measurements correlate to off‐line chemical analysis. In the second application,operandospectroelectrochemical measurements are used to characterize a redox‐active catechol‐based hydrogel film: it is shown that electron transfer into/from the film correlates to the molecular switching of the film's redox state with the film's absorbance increasing upon oxidation and the film's fluorescence increasing upon reduction. In the final example, a synthetic biofilm containing redox‐responsiveE. coliis electro‐assembled: it is shown that gene expression can be induced under reducing conditions (via reductive H₂O₂generation) or oxidative conditions (via oxidation of a phenolic redox‐signaling molecule). Overall, this work demonstrates that 3D printing allows the fabrication of bespoke electrochemical devices that can accelerate the understanding of redox‐based phenomena in biology and enable the detection/characterization redox activities in technology. 

We developed an innovative 3D printed casing that incorporates a lateral-flow immunoassay, dehydrated signal enhancement reagents, and a sealed buffer chamber. With only the push of a button for signal enhancement, our device detected the SARS-CoV-2 N-protein in 40 min at concentrations as low as 0.1 ng mL −1 in undiluted serum. 

Abstract We introduce new analysis methods for studying the star cluster formation processes in Orion A, especially examining the scenario of a cloud–cloud collision. We utilize the CARMA–NRO Orion survey 13CO (1–0) data to compare molecular gas to the properties of young stellar objects from the SDSS III IN-SYNC survey. We show that the increase of $$v_{\rm {}^{13}CO} - v_{\rm YSO}$$ and Σ scatter of older YSOs can be signals of cloud–cloud collision. SOFIA-upGREAT 158 μm [C ii] archival data toward the northern part of Orion A are also compared to the 13CO data to test whether the position and velocity offsets between the emission from these two transitions resemble those predicted by a cloud–cloud collision model. We find that the northern part of Orion A, including regions ONC-OMC-1, OMC-2, OMC-3, and OMC-4, shows qualitative agreements with the cloud–cloud collision scenario, while in one of the southern regions, NGC 1999, there is no indication of such a process in causing the birth of new stars. On the other hand, another southern cluster, L 1641 N, shows slight tendencies of cloud–cloud collision. Overall, our results support the cloud–cloud collision process as being an important mechanism for star cluster formation in Orion A.