A5044785384- Monoclonal and Polyclonal Antibodies Research
- Glycosylation and Glycoproteins Research
- Genetics, Bioinformatics, and Biomedical Research
- CAR-T cell therapy research
- CRISPR and Genetic Engineering
- Peptidase Inhibition and Analysis
- Bacterial Genetics and Biotechnology
- Protein Structure and Dynamics
- Ubiquitin and proteasome pathways
- Enzyme Structure and Function
- HER2/EGFR in Cancer Research
- Cancer Immunotherapy and Biomarkers
- RNA and protein synthesis mechanisms
- Biomedical and Engineering Education
- Radiopharmaceutical Chemistry and Applications
- Antibiotic Resistance in Bacteria
- Galectins and Cancer Biology
- Microbial Metabolic Engineering and Bioproduction
- Immune Cell Function and Interaction
- Protein purification and stability
- Photochromic and Fluorescence Chemistry
- Photosynthetic Processes and Mechanisms
- Photoreceptor and optogenetics research
- Coenzyme Q10 studies and effects
- Chronic Myeloid Leukemia Treatments
Pfizer (United States)
2021
University of North Carolina at Chapel Hill
2010-2014
Johns Hopkins University
2003-2012
Significance Photoactivatable proteins are powerful tools for studying biological processes. Light-induced dimers especially useful because they can be turned on and off with high spatial temporal resolution in living systems, allowing control of protein localization activity. Here, we develop apply methods identifying mutations that improve the effectiveness a light-induced dimer. The engineered switch is modular, used most organisms, has more than 50-fold change binding affinity upon light...
We describe an iterative approach for creating protein switches involving the in vitro recombination of two nonhomologous genes. demonstrate this by recombining genes coding TEM1 β-lactamase (BLA) and Escherichia coli maltose binding (MBP) to create a family MBP–BLA hybrids which is positive or negative effector β-lactam hydrolysis. Some these were effectively “on–off” nature, with altering catalytic activity as much 600-fold. The ability confer effector-dependent growth/no growth phenotype...
Computational algorithms for protein design can sample large regions of sequence space, but suffer from undersampling conformational space and energy function inaccuracies. Experimental screening combinatorial libraries avoids the need accurate functions, has limited access to vast amounts space. Here, we test if these two traditionally alternative, potentially complementary approaches be combined a variant ubiquitin-ligase E6AP that will bind nonnatural partner, NEDD8-conjugating enzyme...
In its basal state, KEAP1 binds the transcription factor NRF2 (Kd = 5 nM) and promotes degradation by ubiquitylation. Changes in redox environment lead to modification of key cysteines within KEAP1, resulting protein accumulation genes important for restoring cellular state. Using phage display a computational loop grafting protocol, we engineered monobody (R1) that is potent competitive inhibitor KEAP1–NRF2 interaction. R1 bound with Kd 300 pM human cells freed from activation promoter....
Generating diverse protein libraries that contain improved variants at a sufficiently high frequency is critical for improving the properties of proteins using directed evolution. Many studies have illustrated how random mutagenesis, cassette DNA shuffling and similar approaches are effective diversity generating methods Very few explored circular permutation, intramolecular relocation N- C-termini protein, as diversity-generating step We subjected library permutations TEM-1 β-lactamase to...
We demonstrate that S1 nuclease converts supercoiled plasmid DNA to unit-length, linear dsDNA through the creation of a single, double-stranded break in molecule. These breaks occur not only origin replication near inverted repeats but also at wide variety locations throughout plasmid. exhibits this activity under conditions typically employed for nuclease's single-stranded activity. Thus, digestion DNA, unlike analogous with DNaseI, effectively halts after first break. This property makes...
We report here the discovery and optimization of a novel T cell retargeting anti-GUCY2C x anti-CD3ε bispecific antibody for treatment solid tumors. Using combination hybridoma, phage display rational design protein engineering, we have developed fully humanized manufacturable CD3 that demonstrates favorable pharmacokinetic properties potent in vivo efficacy. Anti-GUCY2C antibodies derived from mouse hybridomas were first into well-behaved human variable region frameworks with full retention...
ABSTRACT We computationally designed a de novo protein–protein interaction between wild‐type ubiquitin and redesigned scaffold. Our strategy was to incorporate zinc at the interface promote affinity orientation specificity. A large set of monomeric scaffold surfaces were engineered with three‐residue coordination sites, residue H68 docked open site complete tetrahedral site. This single bond intended as hotspot polar for binding, surrounding residues on optimized primarily hydrophobic using...
Despite the success of programmed cell death-1 (PD-1) and PD-1 ligand (PD-L1) inhibitors in treating solid tumors, only a proportion patients respond. Here, we describe first-in-class bifunctional therapeutic molecule, STAR0602, that comprises an antibody targeting germline Vβ6 Vβ10 T receptors (TCRs) fused to human interleukin-2 (IL-2) simultaneously engages nonclonal mode TCR activation with costimulation promote expansion αβ subsets expressing distinct variable β (Vβ) chains. In solution,...
Abstract Background We have previously shown that novel bifunctional “STAR” antibody-fusion molecules comprising germline β chain TCR-targeting antibodies fused to costimulatory cytokines co-engage the TCR and T cell cytokine receptors, thereby promoting activation expansion of subsets cells expressing distinct variable (Vβ) TCRs. Here we describe a new class engager (TriSTAR) utilizes tri-specific format STAR antibody-IL-2 fusion with an additional Fab arm targeting tumor-associated...
Originally published in PROTEINS: Structure, Function, and Bioinformatics 2013;81(7):1245–1255 (DOI: 10.1002/prot.24280) Due to a print error the originally version of this article, author's name is incorrect as published. The correct spelling Ramesh K. Jha.
<h3>Background</h3> Limitations with agents that enhance endogenous T cell responses to cancer, particularly in solid tumors, supports the study of alternative approaches. Directly targeting variable (V) regions receptor (TCR) is a novel approach inducing activation. STAR0602 bispecific antibody-fusion molecule selectively activates and expands subset human aβ cells expressing germline-encoded Vb6 Vb10 TCRs are enriched tumor infiltrating lymphocytes. simultaneously engages novel, non-clonal...