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10.1016/j.celrep.2020.108322 http://scihub22266oqcxt.onion/10.1016/j.celrep.2020.108322 33091382!7550166!33091382     free     free     free Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 267.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 267.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 267.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 267.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 267.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 267.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Cell+Rep 2020 ; 33 (4): 108322 Nephropedia Template TP {{tp|p=33091382|t=2020. Structure-Based Design with Tag-Based Purification and In-Process Biotinylation Enable Streamlined Development of SARS-CoV-2 Spike Molecular Probes |pdf=|usr=}}{{33091382}} gab.com Text Structure-Based Design with Tag-Based Purification and In-Process Biotinylation Enable Streamlined Development of SARS-CoV-2 Spike Molecular Probes JO: Cell Rep http://www.kidney.de/pget.php?&tw=1&pmid=33091382 #FOAvip Biotin-labeled molecular probes, comprising specific regions of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike, would be helpful in the isolation and characterization of antibodies targeting this recently emerged pathogen. Here, we design constructs incorporating an N-terminal purification tag, a site-specific protease-cleavage site, the probe region of interest, and a C-terminal sequence targeted by biotin ligase. Probe regions include full-length spike ectodomain as well as various subregions, and we also design mutants that eliminate recognition of the angiotensin-converting enzyme 2 (ACE2) receptor. Yields of biotin-labeled probes from transient transfection range from approximately 0.5 mg/L for the complete ectodomain to >5 mg/L for several subregions. Probes are characterized for antigenicity and ACE2 recognition, and the structure of the spike ectodomain probe is determined by cryoelectron microscopy. We also characterize antibody-binding specificities and cell-sorting capabilities of the biotinylated probes. Altogether, structure-based design coupled to efficient purification and biotinylation processes can thus enable streamlined development of SARS-CoV-2 spike ectodomain probes. Twit Text FOAVip Structure-Based Design with Tag-Based Purification and In-Process Biotinylation Enable Streamlined Development of SARS-CoV-2 Spike Molecular Probes ????Cell Rep http://www.kidney.de/pget.php?&tw=1&pmid=33091382 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=33091382 #FOAvip English Wikipedia <ref>{{Cite pmid|33091382}}</ref> Structure-Based Design with Tag-Based Purification and In-Process Biotinylation Enable Streamlined Development of SARS-CoV-2 Spike Molecular Probes #MMPMID33091382 Zhou T; Teng IT; Olia AS; Cerutti G; Gorman J; Nazzari A; Shi W; Tsybovsky Y; Wang L; Wang S; Zhang B; Zhang Y; Katsamba PS; Petrova Y; Banach BB; Fahad AS; Liu L; Lopez Acevedo SN; Madan B; Oliveira de Souza M; Pan X; Wang P; Wolfe JR; Yin M; Ho DD; Phung E; DiPiazza A; Chang LA; Abiona OM; Corbett KS; DeKosky BJ; Graham BS; Mascola JR; Misasi J; Ruckwardt T; Sullivan NJ; Shapiro L; Kwong PD Cell Rep 2020[Oct]; 33 (4): 108322 PMID33091382show ga Biotin-labeled molecular probes, comprising specific regions of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike, would be helpful in the isolation and characterization of antibodies targeting this recently emerged pathogen. Here, we design constructs incorporating an N-terminal purification tag, a site-specific protease-cleavage site, the probe region of interest, and a C-terminal sequence targeted by biotin ligase. Probe regions include full-length spike ectodomain as well as various subregions, and we also design mutants that eliminate recognition of the angiotensin-converting enzyme 2 (ACE2) receptor. Yields of biotin-labeled probes from transient transfection range from approximately 0.5 mg/L for the complete ectodomain to >5 mg/L for several subregions. Probes are characterized for antigenicity and ACE2 recognition, and the structure of the spike ectodomain probe is determined by cryoelectron microscopy. We also characterize antibody-binding specificities and cell-sorting capabilities of the biotinylated probes. Altogether, structure-based design coupled to efficient purification and biotinylation processes can thus enable streamlined development of SARS-CoV-2 spike ectodomain probes. |Angiotensin-Converting Enzyme 2[MESH] |Antibodies, Neutralizing/*immunology[MESH] |Antibodies, Viral/*immunology[MESH] |Antibody Specificity/immunology[MESH] |Binding Sites, Antibody/immunology[MESH] |Biotinylation[MESH] |COVID-19[MESH] |Coronavirus Infections/*immunology[MESH] |Cryoelectron Microscopy[MESH] |Humans[MESH] |Molecular Probes/*immunology[MESH] |Pandemics[MESH] |Peptidyl-Dipeptidase A/metabolism[MESH] |Pneumonia, Viral/*immunology[MESH] |Receptors, Virus/metabolism[MESH]Structure-Based Design with Tag-Based Purification and In-Process Biot(epmc).pdf DeepDyve Pubget Overpricing