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Creators/Authors contains: "López, Gabriel P"

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  1. Abstract Extraction of nucleic acids (NAs) is critical for many methods in molecular biology and bioanalytical chemistry. NA extraction has been extensively studied and optimized for a wide range of applications and its importance to society has significantly increased. The COVID-19 pandemic highlighted the importance of early and efficient NA testing, for which NA extraction is a critical analytical step prior to the detection by methods like polymerase chain reaction. This study explores simple, new approaches to extraction using engineered smart nanomaterials, namely NA-binding, intrinsically disordered proteins (IDPs), that undergo triggered liquid–liquid phase separation (LLPS). Two types of NA-binding IDPs are studied, both based on genetically engineered elastin-like polypeptides (ELPs), model IDPs that exhibit a lower critical solution temperature in water and can be designed to exhibit LLPS at desired temperatures in a variety of biological solutions. We show that ELP fusion proteins with natural NA-binding domains can be used to extract DNA and RNA from physiologically relevant solutions. We further show that LLPS of pH responsive ELPs that incorporate histidine in their sequences can be used for both binding, extraction and release of NAs from biological solutions, and can be used to detect SARS-CoV-2 RNA in samples from COVID-positive patients. 
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  2. Most biosensing techniques require complex processing steps that generate prolonged workflows and introduce potential points of error. Here, we report an acoustic pipette to purify and label biomarkers in 70 minutes. A key aspect of this technology is the use of functional negative acoustic contrast particles (fNACPs), which display biorecognition motifs for the specific capture of biomarkers from whole blood. Because of their large size and compressibility, the fNACPs robustly trap along the pressure antinodes of a standing wave and separate from blood components in under 60 seconds with >99% efficiency. fNACPs are subsequently fluorescently labeled in the pipette and are analyzed by both a custom, portable fluorimeter and flow cytometer. We demonstrate the detection of anti-ovalbumin antibodies from blood at picomolar levels (35 to 60 pM) with integrated controls showing minimal nonspecific adsorption. Overall, this system offers a simple and versatile approach for the rapid, sensitive, and specific capture of biomolecules. 
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  3. Employing concepts from physics, chemistry and bioengineering, 'learning-by-building' approaches are becoming increasingly popular in the life sciences, especially with researchers who are attempting to engineer cellular life from scratch. The SynCell2020/21 conference brought together researchers from different disciplines to highlight progress in this field, including areas where synthetic cells are having socioeconomic and technological impact. Conference participants also identified the challenges involved in designing, manipulating and creating synthetic cells with hierarchical organization and function. A key conclusion is the need to build an international and interdisciplinary research community through enhanced communication, resource-sharing, and educational initiatives. 
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