A5087304853- SARS-CoV-2 and COVID-19 Research
- Bacteriophages and microbial interactions
- Monoclonal and Polyclonal Antibodies Research
- Advanced biosensing and bioanalysis techniques
- RNA and protein synthesis mechanisms
- Bacterial Genetics and Biotechnology
- SARS-CoV-2 detection and testing
- Biosensors and Analytical Detection
- Immunotherapy and Immune Responses
- Protein purification and stability
- Microbial Metabolic Engineering and Bioproduction
- COVID-19 Clinical Research Studies
- Glycosylation and Glycoproteins Research
- Genomics and Phylogenetic Studies
- CAR-T cell therapy research
- Transgenic Plants and Applications
- Biochemical and Structural Characterization
- Immune cells in cancer
- Receptor Mechanisms and Signaling
- Extracellular vesicles in disease
- DNA and Nucleic Acid Chemistry
- Virus-based gene therapy research
- Biological Stains and Phytochemicals
- Animal Virus Infections Studies
- Click Chemistry and Applications
University of California, San Francisco
2020-2025
University of California, Berkeley
2012-2018
National Institute of Standards and Technology
2016-2017
Stanford University
2017
The University of Texas at Austin
1967
Significance During the ongoing COVID-19 pandemic, protein engineering offers a rapid and powerful approach for building therapeutics to treat severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections. We use computational design, affinity maturation, fusion dimerization domains engineer “receptor traps” based on wild-type angiotensin-converting enzyme II (ACE2), target viral spike-mediated SARS-CoV-2 entry into cells. The optimized ACE2 receptor traps neutralize authentic...
Our understanding of translation underpins our capacity to engineer living systems. The canonical start codon (AUG) and a few near-cognates (GUG, UUG) are considered as the 'start codons' for initiation in Escherichia coli. Translation is typically not thought initiate from 61 remaining codons. Here, we quantified green fluorescent protein nanoluciferase E. coli all 64 triplet codons across range DNA copy number. We detected synthesis above measurement background 47 non-canonical ranged...
The encapsulation of enzymes in nanometer-sized compartments has the potential to enhance and control enzymatic activity, both vivo vitro. Despite this potential, there is little quantitative data on effect a well-defined compartment under varying conditions. To gain more insight into these effects, we have characterized two improved methods for heterologous molecules inside bacteriophage MS2 viral capsids. First, attaching DNA oligomers molecule interest incubating it with coat protein...
As severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to spread around the world, there is an urgent need for new assay formats characterize humoral response infection. Here, we present efficient, competitive serological that can simultaneously determine individual's seroreactivity against SARS-CoV-2 Spike protein and proportion of anti-Spike antibodies block interaction with human angiotensin-converting enzyme (ACE2) required viral entry. In this approach based on use...
Measuring virus in biofluids is complicated by confounding biomolecules coisolated with viral nucleic acids. To address this, we developed an affinity-based microfluidic device for specific capture of intact severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Our approach used engineered angiotensin-converting enzyme to from plasma and other complex biofluids. leverages a staggered herringbone pattern, nanoparticle surface coating, processing conditions achieve detection as few 3...
The placement of fluorophores in close proximity to metal nanoparticle surfaces is proposed enhance several photophysical properties the dyes, potentially leading improved quantum yields and decreased photobleaching. It difficult practice, however, establish maintain nanoscale distances that are required maximize these effects. type metal, size, shape nanoparticle, physical distance separating from organic dye, spectral fluorophore itself all influence yield lifetime. This results a complex...
Chemoenzymatic modification of proteins is an attractive option to create highly specific conjugates for therapeutics, diagnostics, or materials under gentle biological conditions. However, these methods often suffer from expensive specialized substrates, bulky fusion tags, low yields, and extra purification steps achieve the desired conjugate. Staphylococcus aureus sortase A its engineered variants are used attach oligoglycine derivatives C-terminus expressed with a minimal LPXTG tag. This...
Nature uses protein compartmentalization to great effect for control over enzymatic pathways, and the strategy has promise synthetic biology. In particular, encapsulation in nanometer-sized containers create nanoreactors potential elicit interesting, unexplored effects resulting from deviations well-understood bulk processes. Self-assembled shells are especially desirable their uniform structures ease of perturbation through genetic mutation. Here, we use MS2 capsid, a well-defined porous 27...
Tumor-infiltrating CD8+ T cells mediate antitumor immune responses. However, the mechanisms by which remain poised to kill cancer despite expressing high levels of inhibitory receptors are unknown. Here, we report that layilin, a C-type lectin domain–containing membrane glycoprotein, is selectively expressed on highly activated, clonally expanded, but phenotypically exhausted in human melanoma. Lineage-specific deletion layilin murine reduced their accumulation tumors and increased tumor...
Characterization of cell surface proteome differences between cancer and healthy cells is a valuable approach for the identification novel diagnostic therapeutic targets. However, selective sampling proteins proteomics requires large samples (>10e6 cells) long labeling times. These limitations preclude analysis material-limited biological or capture rapid proteomic changes. Here, we present two approaches to tether exogenous peroxidases (APEX2 HRP) directly cells, enabling rapid, small-scale...
Virus-like particles are used to encapsulate drugs, imaging agents, enzymes, and other biologically active molecules in order enhance their function. However, the size of most virus-like is inflexible, precluding design appropriately sized containers for different applications. Here, we describe a chromatographic selection particle assembly. Using this selection, identified single amino acid substitution coat protein bacteriophage MS2 that mediates uniform switch geometry from T = 3 1...
Abstract Yeast surface display, a well‐established technology for protein analysis and engineering, involves expressing of interest as genetic fusion to either the N‐ or C‐terminus yeast Aga2p mating protein. Historically, yeast‐displayed variants are flanked by peptide epitope tags that enable flow cytometric measurement construct expression using fluorescent primary secondary antibodies. Here, we built upon this develop new display strategy comprises two different proteins Aga2p, one...
An essential mechanism for SARS-CoV-1 and -2 infection begins with the viral spike protein binding to human receptor angiotensin-converting enzyme II (ACE2). Here we describe a stepwise engineering approach generate set of affinity optimized, enzymatically inactivated ACE2 variants that potently block SARS-CoV-2 cells. These optimized traps tightly bind domain (RBD) prevent entry into host We first computationally designed ACE2-RBD interface using two-stage flexible backbone design process...
It has been shown' previously that the corrected sedimentation coefficients (820 W) of ribosomes from 25 species fungi and bacteria, respec- tively, range 76.5 to 84.5 Svedberg units (S) 63.8 72.1S, difference between arithmetic means, 81.3 68.4S, these two frequency distributions is significant.Since other eucaryotic micro- organisms, plants, animals also belong 80S class,2 it appears 70S are characteristic bacteria.Therefore, becomes interest determine whether this uniqueness reflected in...
ABSTRACT As SARS-CoV-2 continues to spread around the world, there is an urgent need for new assay formats characterize humoral response infection. Convalescent serum being used treatment and isolation of patient-derived antibodies. However, currently not a simple means estimate bulk neutralizing capability. Here we present efficient competitive serological that can simultaneously determine individual’s seropositivity against Spike protein capacity anti-Spike antibodies block interaction...
SUMMARY Current serology tests for SARS-CoV-2 antibodies mainly take the form of enzyme-linked immunosorbent assays or lateral flow assays, with former being laborious and latter expensive often lacking sufficient sensitivity scalability. Here we present development validation a rapid, low-cost solution-based assay to detect in serum, plasma, whole blood, saliva, using rationally designed split luciferase antibody biosensors (spLUC). This new assay, which generates quantitative results as...
New epitopes for immune recognition provide the basis of anticancer immunity. Due to high concentration extracellular adenosine triphosphate in tumor microenvironment, we hypothesized that kinases (ectokinases) could have dysregulated activity and introduce aberrant phosphorylation sites on cell surface proteins. We engineered a cell-tethered version kinase CK2α, demonstrated it was active cells under tumor-relevant conditions, profiled its substrate scope using chemoproteomic workflow. then...
ABSTRACT Our understanding of translation is one cornerstone molecular biology that underpins our capacity to engineer living matter. The canonical start codon (AUG) and a few near-cognates (GUG, UUG) are typically considered as the “start codons” for initiation in Escherichia coli ( E. ). Translation not thought initiate from 61 remaining codons. Here, we systematically quantified all 64 triplet We detected protein synthesis above background initiating at least 46 initiated these...