BIOSENSING Biosensing International Research and Development
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Mass Spectrometry of Self-Assembled Monolayers: a New Tool for Molecular Surface Science
REVIEW Mass Spectrometry of Self-Assembled Monolayers: A New Tool for Molecular Surface Science Milan Mrksich* Department of Chemistry and Howard Hughes Medical Institute, The University of Chicago, 929 East 57th Street, Chicago, Illinois 60521 he scientific and engineering disci- T plines have developed sophisti- ABSTRACT Most reactions can be performed in solution and on a surface, yet the challenges faced in applying cated toolboxes for making mat- known reactions or in developing entirely new reactions for modifying surfaces remain formidable. The products ter—including examples from organic of many reactions performed in solution can be characterized in minutes, and even products having complex chemistry to create small molecules with structures can be characterized in hours. When performed on surfaces, even the most basic reactions require a virtually unlimited complexity, molecular bi- ology to prepare macromolecules with a substantial effort—requiring several weeks—to characterize the yields and structures of the products. This precise arrangement of amino acids and a contrast stems from the lack of convenient analytical tools that provide rapid information on the structures of defined tertiary structure, and lithographic molecules attached to a surface. This review describes recent work that has established mass spectrometry as a methods to fashion inorganic and metallic powerful method for developing and characterizing a broad range of chemical reactions of molecules attached to materials into integrated circuits and de- self-assembled monolayers of alkanethiolates on gold. The SAMDI-TOF mass spectrometry technique will enable a vices. Each of these approaches is based on next generation of applications of molecularly defined surfaces to problems in chemistry and biology. -
Peptide Arrays: Development and Application Lindsey C
Review Cite This: Anal. Chem. 2018, 90, 266−282 pubs.acs.org/ac Peptide Arrays: Development and Application Lindsey C. Szymczak,† Hsin-Yu Kuo,‡ and Milan Mrksich*,†,‡,§ § Institute of Chemical Biology and Nanomedicine, Hunan University, Changsha 410082, China † Department of Chemistry, Northwestern University, Evanston, Illinois 60208, United States ‡ Department of Biomedical Engineering, Northwestern University, Evanston, Illinois 60208, United States ■ CONTENTS the immobilization of presynthesized DNA or the in situ synthesis of DNA directly on the substrate, and by 1995 were Technical Factors 267 using the arrays to profile gene expression.4 In 1996, DNA Peptide Synthesis 267 arrays were used to profile the expression of as many as 1000 Solid Support and Surface Functionalization 268 genes5 and soon after, the first whole-genome microarray of Immobilization Methods 269 yeast was reported6 and a new high-density random array was Physical Immobilization 270 7 Chemical Immobilization 270 presented. Now, 25 years later, commercially available arrays contain several million oligonucleotides and are routine tools in Biological Immobilization 271 8,9 Patterning Methods 271 the laboratory. Oligonucleotide arrays have been used in Detection Methods 272 clinical applications to better understand viruses and human disease, for genetic screening, and for the implementation of Current Approaches and Commercialization 273 10,11 Applications 276 personalized medicine. The rapid pace of technical Summary 279 advancement of the early arrays, which performed rather Author Information 279 poorly, has led to the robust and inexpensive arrays currently in Corresponding Author 279 use. By analogy, peptide arrays are still at the early stage of ORCID 279 development and can be expected to see improvements that Notes 279 result in an important, if somewhat less ubiquitous, technology Biographies 279 for the life sciences. -
Synthesis, Characterization, and Simulation of Four-Armed Megamolecules Shengwang Zhou, Peng He, Sonali Dhindwal, Valerie L
pubs.acs.org/Biomac Article Synthesis, Characterization, and Simulation of Four-Armed Megamolecules Shengwang Zhou, Peng He, Sonali Dhindwal, Valerie L. Grum-Tokars, Ying Li, Kelly Parker, Justin A. Modica, Reiner Bleher, Roberto dos Reis, Joshua Zuchniarz, Vinayak P. Dravid, Gregory A. Voth, Benoît Roux, and Milan Mrksich* Cite This: https://doi.org/10.1021/acs.biomac.1c00118 Read Online ACCESS Metrics & More Article Recommendations *sı Supporting Information ABSTRACT: This paper describes the synthesis, characterization, and modeling of a series of molecules having four protein domains attached to a central core. The molecules were assembled with the “megamolecule” strategy, wherein enzymes react with their covalent inhibitors that are substituted on a linker. Three linkers were synthesized, where each had four oligo(ethylene glycol)- based arms terminated in a para-nitrophenyl phosphonate group that is a covalent inhibitor for cutinase. This enzyme is a serine hydrolase and reacts efficiently with the phosphonate to give a new ester linkage at the Ser-120 residue in the active site of the enzyme. Negative-stain transmission electron microscopy (TEM) images confirmed the architecture of the four-armed megamolecules. These cutinase tetramers were also characterized by X-ray crystallography, which confirmed the active-site serine-phosphonate linkage by electron-density maps. Molecular dynamics simulations of the tetracutinase megamolecules using three different force field setups were performed and compared with the TEM observations. Using the Amberff99SB-disp + pH7 force field, the two-dimensional projection distances of the megamolecules were found to agree with the measured dimensions from TEM. The study described here, which combines high-resolution characterization with molecular dynamics simulations, will lead to a comprehensive understanding of the molecular structures and dynamics for this new class of molecules. -
Substrates for Cell Adhesion Prepared Via Active Site-Directed Immobilization of a Protein Domain
1026 Langmuir 2004, 20, 1026-1030 Substrates for Cell Adhesion Prepared via Active Site-Directed Immobilization of a Protein Domain William L. Murphy,†,§ Kwesi O. Mercurius,†,§ Shohei Koide,‡ and Milan Mrksich*,† University of Chicago, Department of Chemistry and Institute for Biophysical Dynamics, and Department of Biochemistry and Molecular Biology, Chicago, Illinois 60637 Received September 16, 2003. In Final Form: December 5, 2003 Recent studies in basic cell biology and bioengineering call for model substrates that present active proteins, with control over protein density, pattern, and orientation, to more directly mimic the natural extracellular matrix. Herein we demonstrate a strategy for controlled, irreversible immobilization of a cell adhesion protein domain onto an otherwise bioinert substrate with well-defined protein orientation and density. Our approach uses a tri(ethylene glycol)-terminated self-assembled monolayer presenting a phosphonate ligand that is used to immobilize an engineered fusion protein. One component of the fusion protein, the 22 kDa serine esterase cutinase, reacts with the surface-bound ligand to form a site-specific covalent adduct, and the second component of the fusion protein is therefore immobilized on the surface. Here we use this approach to immobilize an engineered version of the 12 kDa 10th domain of fibronectin III (FnIII10Eng) to direct cell adhesion. Substrates presenting this protein mediated rapid attachment and spreading of Swiss 3T3 fibroblasts, while substrates presenting cutinase or the phosphonate ligand alone did not support cell attachment. In addition, we used Chinese hamster ovary cells engineered to express specific integrin receptors to show that FnIII10Eng interacts with multiple integrin cell surface receptors, including human Rv- and R5-containing integrins. -
2014 Perspectives
CHICAGO BIOMEDICAL CONSORTIUM The University of Chicago University of Illinois at Chicago Northwestern University 2014 Perspectives NORTHWESTERN UNIVERSITY EVANSTON CAMPUS NORTHWESTERN UNIVERSITY CHICAGO CAMPUS UNIVERSITY OF ILLINOIS AT CHICAGO THE UNIVERSITY OF CHICAGO The CBC is funded by the Searle Funds at The Chicago Community Trust CBC Mission The mission of the Chicago Biomedical Consortium is to stimulate collaboration among scientists at Northwestern University (NU), the University of Chicago (UChicago), and the University of Illinois at Chicago (UIC) that will transform research at the frontiers of biomedicine. The CBC works to: t Stimulate research and education that bridge institutional boundaries t Enable collaborative and interdisciplinary research that is beyond the range of a single institution t Recruit and retain a strong cadre of biomedical leaders and researchers in Chicago t Promote the development of the biomedical industry in Chicago t Execute a plan capable of improving the health of citizens of Chicago and beyond CBC Leadership and Staff Brian Kay, PhD, Scientific Director Professor, Department of Biological Sciences, University of Illinois at Chicago Shohei Koide, PhD, Scientific Director Professor, Biochemistry and Molecular Biology, The University of Chicago Richard Morimoto, PhD, Scientific Director Bill and Gayle Cook Professor of Biology, Director, Rice Institute for Biomedical Research, Department of Molecular Biosciences, Northwestern University Kathryn Stallcup, PhD, Executive Director Karen Snapp, DDS, PhD, Senior Associate Director Kimberly Corn, Associate Director, Business Operations and Finance Jola Glotzer, MD, Communications Director Corinna Kitcharoen, MBA, Program Coordinator Front Cover Credits Images of fluorescently-labeled cells overlaid on a Google earth Photos: Peter Barreras, Howard Hughes Medical Institute (Chuan (©2013 Google) image of the Chicago area.