A5068292183- Monoclonal and Polyclonal Antibodies Research
- Cellular Mechanics and Interactions
- Protein purification and stability
- Cardiomyopathy and Myosin Studies
- Glycosylation and Glycoproteins Research
- Muscle Physiology and Disorders
- Microtubule and mitosis dynamics
- CAR-T cell therapy research
- T-cell and B-cell Immunology
- Force Microscopy Techniques and Applications
- Viral Infectious Diseases and Gene Expression in Insects
- NF-κB Signaling Pathways
- Cellular transport and secretion
- Nuclear Structure and Function
- Chromosomal and Genetic Variations
- Immune Response and Inflammation
- Immune Cell Function and Interaction
- Advanced Fluorescence Microscopy Techniques
- COVID-19 Clinical Research Studies
- Erythrocyte Function and Pathophysiology
- Bacteriophages and microbial interactions
- Reproductive System and Pregnancy
- Bacterial biofilms and quorum sensing
- Skin and Cellular Biology Research
- Connexins and lens biology
Johnson & Johnson (United States)
2023-2025
Janssen (United States)
2017-2024
Springhouse
2022
University of Pennsylvania
2013-2018
University of Washington
2018
National Heart Lung and Blood Institute
2011
National Institutes of Health
2010
New York University
2010
Saint Joseph's University
2008
Myosins adjust their power outputs in response to mechanical loads an isoform-dependent manner, resulting ability dynamically adapt a range of motile challenges. Here, we reveal the structural basis for force-sensing based on near-atomic resolution structures one rigor and two ADP-bound states myosin-IB (myo1b) bound actin, determined by cryo-electron microscopy. The are separated 25° rotation lever. lever first ADP state is rotated toward pointed end actin filament forms previously...
Treating and preventing infections by antimicrobial-resistant bacterial pathogens is a worldwide problem. Pathogens such as Staphylococcus aureus produce an array of virulence determinants, making it difficult to identify single targets for the development vaccines or monoclonal therapies. We described human-derived anti-S. antibody (mAb)-centyrin fusion protein ("mAbtyrin") that simultaneously multiple adhesins, resists proteolysis protease GluV8, avoids Fc engagement S. IgG-binding...
Single-chain fragment variable (scFv) domains play an important role in antibody-based therapeutic modalities, such as bispecifics, multispecifics and chimeric antigen receptor T cells or natural killer cells. However, scFv exhibit lower stability increased risk of aggregation due to transient dissociation ("breathing") inter-molecular reassociation the two (VL VH). We designed a novel strategy, referred stapling, that introduces disulfide bonds between linker minimize breathing. named...
T cell-redirecting bispecific antibodies (bsAbs) to treat advanced stage solid tumors are gaining interest after recent clinical successes. The immune checkpoint human leukocyte antigen G (HLA-G) is expressed in several tumor types while normal tissues expression limited. Here, we describe JNJ-78306358, a antibody (bsAb) tumors. JNJ-78306358 binds with high affinity the α3 subunit of HLA-G on cancer cells and purposely engineered weaker CD3ε cells. induced potent cell-mediated cytotoxicity...
Significance We report the high-resolution structure of a tension-sensing myosin-Ib. identify striking unique orientation structural elements that position motor’s lever arm. This results in cavity between motor and arm holds 10-residue stretch N-terminal amino acids, region is divergent among myosins. show importance myosin controlling kinetics mechanics motor.
PICK1 is a modular scaffold implicated in synaptic receptor trafficking. It features PDZ domain, BAR and an acidic C-terminal tail (ACT). Analysis by small- angle x-ray scattering suggests structural model that places the receptor-binding site of domain membrane-binding surfaces domains adjacent to each other on concave side banana-shaped dimer. In model, ACT one subunit dimer interacts with subunit, possibly accounting for autoinhibition. Consistently, full-length shows diffuse cytoplasmic...
Capping protein (CP) is a ubiquitously expressed, 62-kDa heterodimer that binds the barbed end of actin filament with ∼0.1 nm affinity to prevent further monomer addition. CARMIL multidomain protein, present from protozoa mammals, CP and important for normal dynamics in vivo. The binding site resides its CAH3 domain (CARMIL homology 3) located at or near protein's C terminus. ∼1 affinity, resulting complex weak capping activity (30–200 nm). Solution assays single-molecule imaging show...
Abstract Generation of bispecific antibodies (BsAbs) having two unique Fab domains requires heterodimerization the heavy chains and pairing each chain with its cognate light chain. An alternative scaffold (Bipod) comprising an scFv a on heterodimeric Fc eliminates possibility mispairing. However, unpredictable levels expression scFv-induced aggregation can complicate purification reduce yield desired Bipod. Here, we describe high-throughput method for generation Bipods based protein A CH1...
Methods to rapidly generate high quality bispecific antibodies (BsAb) having normal half-lives are critical for therapeutic programs. Here, we identify 3 mutations (T307P, L309Q, and Q311R or "TLQ") in the Fc region of human IgG1 which disrupt interaction with protein A while enhancing FcRn. The shown incrementally alter pH at a mAb elutes from affinity resin. BsAb comprised TLQ mutant wild-type can be efficiently separated contaminating parental mAbs by differential elution starting either...
The serum half-life and clearance of therapeutic monoclonal antibodies (mAbs) are critical factors that impact their efficacy optimal dosing regimen. pH-dependent binding an mAb to the neonatal Fc receptor (FcRn) has long been recognized as important determinant its pharmacokinetics. However, FcRn affinity alone is not a reliable predictor half-life, suggesting other biologic or biophysical mechanisms must be accounted for. thermal stability, which reflects unfolding aggregation...
The availability of the complete genome sequence Bdellovibrio bacteriovorus provides an opportunity for investigating genes that play a significant role in predation. Using two independently derived facultatively predatory strains, we have designed method to cultivate and screen transposon insertion mutants 96-well microtiter dishes. Transposon were produced by introducing plasposon pRL27, which carries mini-Tn5. Mutants been screened activity using plates. Seventeen independent nonpredatory...
Bispecific antibodies (bsAbs) combine the antigen specificities of two distinct Abs and demonstrate therapeutic promise based on novel mechanisms action. Among many platforms for creating bsAbs, controlled Fab-arm exchange (cFAE) has proven useful minimal changes to native Ab structure simplicity with which bsAbs can be formed from parental Abs. Despite a published protocol cFAE its widespread use in pharmaceutical industry, reaction mechanism not been determined. Knowledge could lead...
The increased number of bispecific antibodies (BsAb) under therapeutic development has resulted in a need for mouse surrogate BsAbs. Here, we describe one-step method generating highly pure BsAbs suitable vitro and vivo studies. We identify two mutations the IgG2a IgG2b Fc region: one that eliminates protein A binding enhances by 8-fold. show harboring these can be purified from residual parental monoclonal step using affinity chromatography. structural basis effects was analyzed X-ray...
Mucosal immunity is dominated by secretory IgA and IgM, although these are less favorable compared to IgG molecules for therapeutic development. Polymeric IgM actively transported across the epithelial barrier via engagement of polymeric Ig receptor (pIgR), but lack a lumen-targeted active transport mechanism, resulting in poor biodistribution therapeutics mucosal tissues. In this work, we describe discovery characterization single-domain antibodies (VHH) that engage pIgR undergo...
The global health crisis and economic tolls of COVID-19 necessitate a panoply strategies to treat SARS-CoV-2 infection. To date, few treatment options exist, although neutralizing antibodies against the spike glycoprotein have proven be effective. Because infection is initiated at mucosa propagates mainly this site throughout course disease, blocking virus mucosal milieu should However, administration biologics presents substantial challenge. Here, we describe bifunctional molecules...
Monoclonal antibodies (mAbs) have become an important class of therapeutics, particularly in the realm anticancer immunotherapy. While two antigen-binding fragments (Fabs) mAb allow for high-avidity binding to molecular targets, crystallizable fragment (Fc) engages immune effector elements. mAbs IgG are used treatment autoimmune diseases and can elicit antitumor functions not only by several mechanisms including direct antigen engagement via their Fab arms but also tumors combined with Fc...
Abstract JNJ-78306358 is a first-in-class bispecific antibody (bsAb), engineered using the Zymeworks Azymetric™ platform, to treat advanced stage solid tumors. Human leukocyte antigen G (HLA-G) non-classical major histocompatibility class I molecule with an immune tolerance role at maternal-fetal interface. HLA-G has limited normal tissue expression, mainly detected in placenta and pituitary gland. However, expressed multiple human cancers, potential cancer evasion. Comprehensive...
TL1A (TNFSF15) is a TNF superfamily ligand which can bind the TNFRSF member death receptor 3 (DR3) on T cells and soluble decoy DcR3. Engagement of DR3 CD4+ or CD8+ effector by induces downstream signaling, leading to proliferation an increase in secretion inflammatory cytokines. We designed stable recombinant molecule that (1) displays high monodispersity stability, (2) ability activate vitro vivo, (3) lacks binding DcR3 while retaining functional activity via DR3. Together these results...