A5036613021- Monoclonal and Polyclonal Antibodies Research
- Chemical Synthesis and Analysis
- vaccines and immunoinformatics approaches
- RNA and protein synthesis mechanisms
- Immunotherapy and Immune Responses
- Computational Drug Discovery Methods
- Cervical Cancer and HPV Research
- Glycosylation and Glycoproteins Research
- Microfluidic and Capillary Electrophoresis Applications
- SARS-CoV-2 detection and testing
- Hepatitis B Virus Studies
- Biochemical and Structural Characterization
- SARS-CoV-2 and COVID-19 Research
- Organic Electronics and Photovoltaics
- Inhalation and Respiratory Drug Delivery
- T-cell and B-cell Immunology
- Advanced biosensing and bioanalysis techniques
- Perovskite Materials and Applications
- Cancer-related Molecular Pathways
- Metal-Catalyzed Oxygenation Mechanisms
- Biotin and Related Studies
- Peptidase Inhibition and Analysis
- Metal complexes synthesis and properties
- Antimicrobial Peptides and Activities
- Conducting polymers and applications
Massachusetts Institute of Technology
2017-2023
University of Florida
2013
Fully synthetic whole proteins in reach Solid-phase peptide synthesis of homogeneous peptides longer than about 50 amino acids has been a long-standing challenge because inefficient coupling and side reactions. Hartrampf et al. used an automated chemistry platform to optimize fast-flow were able produce fully single-domain (see the Perspective by Proulx). The targets included proinsulin enzymes such as barnase version HIV-1 protease containing multiple noncanonical acids. Refolded nearly...
Abstract High-diversity genetically-encoded combinatorial libraries (10 8 −10 13 members) are a rich source of peptide-based binding molecules, identified by affinity selection. Synthetic can access broader chemical space, but typically examine only ~ 10 6 compounds screening. Here we show that in-solution selection be interfaced with nano-liquid chromatography-tandem mass spectrometry peptide sequencing to identify binders from fully randomized synthetic members—a 100-fold gain in diversity...
The β-coronavirus SARS-CoV-2 has caused a global pandemic. Affinity reagents targeting the spike protein are of interest for development therapeutics and diagnostics. We used affinity selection–mass spectrometry rapid discovery synthetic high-affinity peptide binders receptor binding domain (RBD) protein. From library screening with 800 million peptides, we identified three sequences nanomolar affinities (dissociation constants Kd = 80–970 nM) RBD selectivity over human serum proteins....
Significance Combinatorial protein libraries—prepared via molecular biology-based approaches—are invaluable tools for engineering. The inclusion of noncanonical amino acids in such libraries is considerable interest. However, at present no approach competes with chemical synthesis terms the variety and number that can be simultaneously incorporated into a molecule. Here, we describe selection from synthetic as strategy enables identification small (∼30 aa), functional variants comprising...
Human papillomavirus (HPV) infections account for nearly all cervical cancer cases, which is the fourth most common in women worldwide. High-risk variants, including HPV16, drive tumorigenesis part by promoting degradation of tumor suppressor p53. This mediated HPV early protein 6 (E6), recruits E3 ubiquitin ligase E6AP and redirects its activity towards ubiquitinating Targeting interaction interface between E6 a promising modality to mitigate HPV-mediated In this study, we designed covalent...
A methodology to achieve high-throughput de novo sequencing of synthetic peptide mixtures is reported. The approach leverages shotgun nanoliquid chromatography coupled with tandem mass spectrometry-based library (up 2000 peptides) as well automated data analysis protocols filter away incorrect assignments, noise, and side-products. For increasing the confidence in results, spectrometry-friendly designs were developed that enabled unambiguous decoding up 600 sequences per hour while...
This work reports an automated affinity selection-mass spectrometry (AS-MS) approach amenable to both de novo peptide binder discovery and maturation of known binders in a high-throughput selective manner.
Reported is a systematic molecular structure–property relationship study to evaluate the consequences of dedicated H-bonding interactions between electron donors on assembly, absorption, charge collection, and performance in small-molecule bulk heterojunction organic photovoltaic devices. Three families branched quaterthiophene donor chromophores have been synthesized with members that share nearly identical electronic optical properties molecularly dispersed state but are either capable or...
Nature has three biopolymers: oligonucleotides, polypeptides, and oligosaccharides. Each biopolymer independent functions, but when needed, they form mixed assemblies for higher-order purposes, as in the case of ribosomal protein synthesis. Rather than forming large complexes to coordinate role different biopolymers, we dovetail amino acids nucleobases into a single low molecular weight precision polyamide polymer. We established efficient chemical synthesis de novo sequencing procedures...
Rapid discovery and development of serum-stable, selective, high affinity peptide-based binders to protein targets are challenging. Angiotensin converting enzyme 2 (ACE2) has recently been identified as a cardiovascular disease biomarker the primary receptor utilized by severe acute respiratory syndrome coronavirus 2. In this study, we report peptidomimetic ACE2 via selection-mass spectrometry (AS-MS). Multiple ACE2-binding peptides (ABP) were selection from canonical noncanonical libraries...
<p>Ribosomes produce most proteins of living cells in seconds. Here we report highly efficient chemistry matched with an automated fast-flow instrument for the direct manufacturing peptide chains up to 164 amino acids over 328 consecutive reactions. The machine is rapid - chain elongation complete hours. We demonstrate utility this approach by chemical synthesis nine different protein that represent enzymes, structural units, and regulatory factors. After purification folding,...
3-Aminotyrosine (NH2Y) has been a useful probe to study the role of redox active tyrosines in enzymes. This report describes properties NH2Y key importance for its application mechanistic studies. By combining tRNA/NH2Y-RS suppression technology with model protein tailored amino acid studies (α3X, X = NH2Y), formal reduction potential NH2Y32(O•/OH) (E°′ 395 ± 7 mV at pH 7.08 0.05) could be determined using film voltammetry. We find that ΔE°′ between and Y32(O•/OH) when measured under...
Significance Erythrocyte-bound antigens can drive immune tolerance in an antigen-specific fashion. Exploiting this phenomenon, we developed a general strategy to promote by engineering peptide and protein bind erythrocytes. Here, showed that fully d -chiral library be selected vivo for the de novo discovery of robust erythrocyte binder, which attached antigens. An administration engineered mitigated inflammatory responses, suggesting generalizability tolerance-induction validating our ligand...
Here, we combined unsupervised machine learning (ML), non-natural amino acids, and affinity-selection mass-spectrometry (AS-MS) for the discovery of ultra-high affinity peptidomimetics that bind to a protein target. Peptides were discovered using AS-MS, encoded diverse representations, decomposed into two-dimensional “maps” chemical space by dimensionality reduction. These maps showed well-defined clusters target-specific binders distinct from remaining included nonspecific nonbinding...
Ribosomes produce most proteins of living cells in seconds. Here we report highly efficient chemistry matched with an automated fast-flow instrument for the direct manufacturing peptide chains up to 164 amino acids over 328 consecutive reactions. The machine is rapid - chain elongation complete hours. We demonstrate utility this approach by chemical synthesis nine different protein that represent enzymes, structural units, and regulatory factors. After purification folding, synthetic...
Abstract Targeting of antigens to erythrocytes can be used selectively mitigate their immunogenicity, but the methods equip a variety cargoes with erythrocyte-targeting properties are limited. Here we identified D-peptide that targets murine and decreases anti-drug antibody responses when conjugated protective antigen from Bacillus anthracis , protein therapeutic interest. The likewise inflammatory anti-ovalbumin (OVA) CD8 + T cell attached peptide derived OVA. To discover this targeting...
Abstract The β-coronavirus SARS-CoV-2 has caused a global pandemic. Affinity reagents targeting the spike protein, most exposed surface structure of virus, are interest for development therapeutics and diagnostics. We used affinity selection-mass spectrometry rapid discovery synthetic high peptide binders receptor binding domain (RBD) protein. From library screening with 800 million peptides, we identified three sequences nanomolar affinities (dissociation constants K d = 80 to 970 nM) RBD...
ABSTRACT Human papillomavirus (HPV) infections account for nearly all cervical cancer cases, which is the fourth most common in women worldwide. High-risk variants, including HPV16, drive tumorigenesis part by promoting degradation of tumor suppressor p53. This mediated HPV early protein 6 (E6), recruits E3 ubiquitin ligase E6AP and redirects its activity towards ubiquitinating Targeting interaction interface between E6 a promising modality to mitigate HPV-mediated In this study, we designed...