A5017661210- Respiratory viral infections research
- Mass Spectrometry Techniques and Applications
- Pneumonia and Respiratory Infections
- Protein Structure and Dynamics
- Chemical Synthesis and Analysis
- Advanced Proteomics Techniques and Applications
- Herpesvirus Infections and Treatments
- Monoclonal and Polyclonal Antibodies Research
- Bacteriophages and microbial interactions
- Analytical Chemistry and Chromatography
- Viral gastroenteritis research and epidemiology
- Toxin Mechanisms and Immunotoxins
- Reproductive tract infections research
- Crystallization and Solubility Studies
- Parvovirus B19 Infection Studies
- X-ray Diffraction in Crystallography
- Metabolomics and Mass Spectrometry Studies
- Urinary Tract Infections Management
- Animal Virus Infections Studies
- RNA and protein synthesis mechanisms
- DNA and Nucleic Acid Chemistry
- SARS-CoV-2 and COVID-19 Research
- Machine Learning in Bioinformatics
- Supramolecular Self-Assembly in Materials
- Biosensors and Analytical Detection
Merck & Co., Inc., Rahway, NJ, USA (United States)
2016-2025
United States Military Academy
2010-2023
Merck (Germany)
2017
Sidney Kimmel Cancer Center
2004
Scripps Research Institute
2000-2002
University of Zurich
1995-2001
University of Konstanz
1995
We report the results of our work to facilitate protein identification using tandem mass spectra and sequence databases. describe a parallel version SEQUEST (SEQUEST-PVM) that is tolerant toward arithmetic exceptions. The changes we effectively separate search processes on slave nodes from each other. Therefore, if one drops out cluster due an error, rest will carry process end. has been widely used for identifications. modifications made code improve its stability effectiveness in...
Significance Congenital human cytomegalovirus (HCMV) infection is an important cause of newborn disability, and developing a vaccine against congenital HCMV top priority. However, despite decades efforts, remains elusive. Previous vaccines lacked antigen called pentameric glycoprotein H (gH) complex, essential for the virus to infect epithelial/endothelial cells, these induced poor neutralizing antibodies. To support unique concept featuring gH we established 45 mAbs from rabbit immunized...
Background: There is an unmet medical need to develop a vaccine targeting endemic coronaviruses. Antigen-specific monoclonal antibodies (mAbs) are crucial for many assays support development. Objective: In this study, we used the HuCal Fab phage display library with diversity of 4.5 × 1010 identify specific spike proteins four coronaviruses: OC43, NL63, 229E, and HKU1. Methods: As proof concept, established newly designed platform using long-read NGS workflow antibody discovery compared...
Respiratory syncytial virus (RSV) results in significant disease infants, young children, and the elderly. Thus, development of an effective vaccine for these populations is a priority.
The stability of a coiled coil or leucine zipper is controlled by hydrophobic interactions and electrostatic forces between the constituent helices. We have designed 30-residue peptide with repeating seven-residue pattern coil, (abcdefg)n, Glu in positions e g each heptad. glutamate side chains prevented folding at pH values above 6 because repulsion across helix dimer interface as well within individual Protonation carboxylates changed conformation from random monomer to dimer. Folding...
Respiratory syncytial virus (RSV) and human metapneumovirus (hMPV) vaccines have been long overdue. Structure-based vaccine design created a new momentum in the last decade, first RSV finally approved older adults pregnant individuals. These are based on recombinant stabilized pre-fusion F glycoproteins administered as soluble proteins. Multimeric antigenic display could markedly improve immunogenicity should be evaluated next generations of vaccines. Here we tested like particles-based...
The dimer interface of a leucine zipper involves hydrophobic as well electrostatic interactions between the component helices. Here we ask how effects and repulsion balance rate folding thermodynamic stability designed dimeric formed by acidic peptide A that contains four repeating sequence units, (abcdefg)4. aliphatic d residues were same in GCN4 but e g positions occupied Glu, which prevented above pH 6 because repulsion. Leucine A2 was protonation side chains with sharp transition...
A fully human monoclonal antibody (CS-D7, IgG1) specific for the iron regulated surface determinant B (IsdB) of Staphylococcus aureus was isolated from Cambridge Antibody Technology (CAT) scFv library. As compared to previously described IsdB murine monoclonals, CS-D7 has a unique, non-overlapping binding site on IsdB, and exhibits increased in vivo activity. The recognizes conformational epitope spanning amino acids 50 285 affinity 340 (± 75) pM IsdB. bound wide variety S. strains, but not...
Herpes zoster (HZ) (shingles) is the clinical manifestation of varicella-zoster virus (VZV) reactivation. HZ typically develops as people age, due to decreased cell-mediated immunity. However, importance antibodies for immunity against prevention remains be understood. The goal this study was examine breadth and functionality VZV-specific after vaccination with a live attenuated vaccine (Zostavax). Direct enumeration antibody-secreting cells (ASCs) via enzyme-linked immunosorbent spot assay...
Infection with Respiratory Syncytial Virus (RSV) causes both upper and lower respiratory tract disease in humans, leading to significant morbidity mortality young children older adults. Currently, there is no licensed vaccine available, therapeutic options are limited. During the infection process, type I viral fusion (F) glycoprotein on surface of RSV particle rearranges from a metastable prefusion conformation highly stable postfusion form. In people naturally infected RSV, most potent...
Human metapneumovirus (hMPV) belongs to the Pneumoviridae family and is closely related respiratory syncytial virus (RSV). The surface fusion (F) glycoprotein mediates viral primary target of neutralizing antibodies against hMPV. Here we report 113 hMPV-F specific monoclonal (mAbs) isolated from memory B cells human donors. We characterize antibodies' germline usage, epitopes, neutralization potencies, binding specificities. find that unlike RSV-F mAbs, antibody responses hMPV F are less...
Surface-embedded glycoproteins, such as the spike protein trimers of coronaviruses MERS, SARS-CoV, and SARS-CoV-2, play a key role in viral function are target antigen for many vaccines. However, their significant glycan heterogeneity poses an analytical challenge. Here, we utilized individual ion mass spectrometry (I
Coiled coils are simple models for studying the association of two polypeptide chains to form a folded protein. Previous work has shown that folding coiled coil can be described by two-state transition between unfolded monomeric peptide and dimer. Here we report thermodynamic activation parameters unfolding unrelated coils: C62GCN4 A2. corresponds 62 C-terminal residues yeast transcription factor GCN4. The forms dimeric through its 33 residues. A2 is designed 30-residue whose induced low pH...
Chlamydia trachomatis is a human pathogen which causes number of pathologies, including genital tract infections in women that can result tubal infertility. Prevention infection and disease control might be achieved through vaccination; however, safe, efficacious cost-effective vaccine against C. remains an unmet medical need. major outer membrane protein (MOMP), β-barrel integral protein, the most abundant antigen bacterium has been evaluated as subunit candidate. Recombinant MOMP (rMOMP)...
Abstract The development of “soft” ionization methods has enabled the mass spectrometric analysis higher‐order structural features proteins. We have applied electrospray spectrometry (ESI‐MS) to number and composition polypeptide chains in homomeric heteromeric leucine zippers. In comparison with other that been used analyze zippers, such as analytical ultracentrifugation, gel chromatography, or electrophoretic band shift assays, ESI‐MS is very fast highly sensitive provides a...
Protein stability in vitro can be influenced either by introduction of mutations or changes the chemical composition solvent. Recently, we have characterized thermodynamic and rate folding engineered dimeric leucine zipper A2, which has a strengthened hydrophobic core [Dürr, E., Jelesarov, I., Bosshard, H. R. (1999) Biochemistry 38, 870−880]. Here report on energetic consequences cavity introduced Leu/Ala substitution at tightly packed interface how addition 30% glycerol affects...
Abstract Coiled coils consist of two or more amphipathic α–helices wrapped around each other to form a superhelical structure stabi–lized at the interhelical interface by hydrophobic residues spaced in repeating 3–4 sequence pattern. Dimeric coiled have been shown often single step reaction which asso–ciation and folding peptide chains are tightly coupled. Here, we ask whether such simple mechanism may also apply formation three–stranded coil. The designed 29–residue LZ16A was previously be...
Abstract Chlamydial major outer membrane protein (MOMP) is the constituent of bacterial pathogen Chlamydia trachomatis. trachomatis Serovars D–K are leading cause genital tract infections which can lead to infertility or ectopic pregnancies. A vaccine against highly desirable but currently not available. MOMP accounts for ~ 60% chlamydial mass and considered be one candidates C. . We report on spectroscopic analysis native D, E, F, J as well muridarum by size exclusion chromatography multi...
Antibody specificity is limited, and different antigens may cross‐react to varying degrees with the same antibody. An example monoclonal antibody 42PF elicited against a 29‐residue random‐coil pep‐tide. cross‐reacts triple‐stranded coiled coil of related amino acid sequence, there was circumstantial evidence for an antibody‐induced opening [Leder, L., Berger, C., Bornhauser, S., Wendt, H., Ackermann, F., Jelesarov, I. & Bosshard, H. R. (1995) Biochemistry 34 , 16509–16518]. To reveal in...
Surface-embedded glycoproteins, such as the spike protein trimers of coronaviruses MERS, SARS-CoV, and SARS-CoV-2 play a key role in viral function are target antigen for many vaccines. However, their significant glycan heterogeneity poses an analytical challenge. Here, we have utilized individual ion mass spectrometry (I2MS), form charge detection (CDMS) that uses commercially available Orbitrap analyzer, to directly produce heterogeneous profiles these three coronavirus under native-like...
Outer membrane vesicles (OMVs) are lipid structures containing various biomolecules in their native environment and spontaneously shed by gram-negative bacteria. OMVs perform several biological functions critical to both bacterial physiology pathogenicity. Scientific research on OMV function biogenesis requires a standardized robust method of isolating these from cultures that reliably provide high-purity OMVs. Herein, we describe an optimized protocol isolate overnight three different...