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KTH Royal Institute of Technology

6 ARTICLES PUBLISHED IN JoVE

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Education

Unraveling Entropic Rate Acceleration Induced by Solvent Dynamics in Membrane Enzymes
Charlotte Kürten 1, Per-Olof Syrén 1
1Science for Life Laboratory, School of Biotechnology, KTH Royal Institute of Technology

Channels for the transportation of water molecules in enzymes influence active site solvation and catalysis. Herein we present a protocol for the engineering of these additional catalytic motifs based on in silico computer modeling and experiments. This will enhance our understanding of the influence of solvent dynamics on enzyme catalysis.

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Bioengineering

Film Extrusion of Crambe abyssinica/Wheat Gluten Blends
Mikael Gällstedt 1, Henrik Pettersson 2, Therese Johansson 2, William R. Newson 3, Eva Johansson 3, Mikael S. Hedenqvist 4
1SIG Combibloc, 2Innventia, 3Department of Plant Breeding, The Swedish University of Agricultural Sciences, 4Fibre and Polymer Technology, KTH Royal Institute of Technology

The side stream from plant oil production of Crambe abyssinica has limited value. The purpose of this study was to find methods for extruding materials based on this side stream, demonstrating that products with a higher value can be produced. The extrudates were found to have promising properties.

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JoVE Journal

Method to Visualize and Analyze Membrane Interacting Proteins by Transmission Electron Microscopy
Ramakrishnan B. Kumar 1, Lin Zhu 2, Hans Hebert 1,2, Caroline Jegerschöld 1,2
1Department of Biosciences and Nutrition, Karolinska Institutet, 2School of Technology and Health, KTH Royal Institute of Technology

Many proteins perform their function when attached to membrane surfaces. The binding of extrinsic proteins on nanodisc membranes can be indirectly imaged by transmission electron microscopy. We show that the characteristic stacking (rouleau) of nanodiscs induced by the negative stain sodium phosphotungstate is prevented by the binding of extrinsic protein.

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Chemistry

Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis
Jorge T. Dias 1, Gustav Svedberg 1, Mats Nystrand 2, Helene Andersson-Svahn 1, Jesper Gantelius 1
1Division of Proteomics and Nanobiotechnology, Science for Life Laboratory, KTH Royal Institute of Technology, 2Global Research and Development, Thermo Fisher Scientific IDD

In this work, a protocol for signal enhancement of nanoprobe-based biosensing is presented. The protocol is based on the reduction of chloroauric acid onto the surface of existing nanoprobes that consist of gold, silver, silica or iron-oxide nanoparticles.

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Developmental Biology

Generation of a Human iPSC-Based Blood-Brain Barrier Chip
Srikanth Jagadeesan 1,2,3, Michael J. Workman 4, Anna Herland 5,6, Clive N. Svendsen 4, Gad D. Vatine 1,2,3
1The Department of Physiology and Cell Biology, Faculty of Health Sciences, Ben-Gurion University of the Negev, 2The Regenerative Medicine and Stem Cell (RMSC) Research Center, Ben-Gurion University of the Negev, 3The Zlotowski Center for Neuroscience, Ben-Gurion University of the Negev, 4The Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, 5Division of Micro and Nanosystems, KTH Royal Institute of Technology, 6AIMES, Department of Neuroscience, Karolinska Institutet

The blood-brain barrier (BBB) is a multicellular neurovascular unit tightly regulating brain homeostasis. By combining human iPSCs and organ-on-chip technologies, we have generated a personalized BBB chip, suitable for disease modeling and CNS drug penetrability predictions. A detailed protocol is described for the generation and operation of the BBB chip.

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Immunology and Infection

Profiling of Surface Protein Epitopes on Viral Particles by Multiplex Dual-Reporter Strategy
Maryam Sahi 1, Sarah Andersson 1, Cecilia Mattson 1, Matilda Dale 1, Sofia Kagiolglou 1, Camilla Hofström 2, Helena Persson 2, Jonas Klingström 3,4, Francesca Chiodi 5, Claudia Fredolini 1
1Affinity Proteomics-Stockholm Unit, SciLifeLab, Division of Affinity Proteomics, Department of Protein Science, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), KTH Royal Institute of Technology, 2Human Antibody Therapeutics Unit, SciLifeLab, Division of Drug Discovery and Development, Department of Protein Science, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), KTH Royal Institute of Technology, 3Center for Infectious Medicine, Department of Medicine Huddinge, Karolinska Institutet, 4Public Health Agency of Sweden, 5Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet

Here, we describe a newly developed multiplex fluorescent immunoassay that uses a dual-reporter flow cytometric system to concurrently detect two unique spike protein epitopes on intact severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral particles that had been captured by angiotensin-converting enzyme-2-coupled magnetic microspheres.

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