A5017305191- Virus-based gene therapy research
- Glycosylation and Glycoproteins Research
- Mass Spectrometry Techniques and Applications
- Advanced Proteomics Techniques and Applications
- CRISPR and Genetic Engineering
- Viral Infectious Diseases and Gene Expression in Insects
- Viral gastroenteritis research and epidemiology
- Protein purification and stability
- Metabolomics and Mass Spectrometry Studies
- Viral Infections and Immunology Research
- RNA and protein synthesis mechanisms
- Urological Disorders and Treatments
- RNA Interference and Gene Delivery
- Molecular Biology Techniques and Applications
- Surgical Simulation and Training
- Cancer, Hypoxia, and Metabolism
- Uterine Myomas and Treatments
- Carbohydrate Chemistry and Synthesis
- Galectins and Cancer Biology
- Molecular spectroscopy and chirality
- Bacteriophages and microbial interactions
- Anatomy and Medical Technology
- Proteoglycans and glycosaminoglycans research
- Monoclonal and Polyclonal Antibodies Research
- Synthesis and Properties of Aromatic Compounds
Pfizer (United States)
2020-2025
Medical University of South Carolina
2013-2020
Northwestern University
2002-2005
Arcadia
1992-1995
Houston Methodist
1995
Methodist Hospital
1995
University of Illinois Chicago
1982
Orthopaedic Research Society
1982
University of California, Los Angeles
1971
A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended improved for direct analysis glycans clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. Formalin-fixed from normal mouse kidney, human pancreatic prostate cancers, a hepatocellular carcinoma tissue microarray were processed by antigen retrieval...
Abstract Adeno-associated viruses (AAVs) are increasingly used as gene therapy vectors. AAVs package their genome in a non-enveloped T = 1 icosahedral capsid of ~3.8 megaDalton, consisting 60 subunits 3 distinct viral proteins (VPs), which vary only N-terminus. While all three VPs play role cell-entry and transduction, precise stoichiometry structural organization the has remained elusive. Here we investigate composition several AAV serotypes by high-resolution native mass spectrometry. Our...
A new matrix assisted laser desorption ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in tissues is described. Application an endoglycosidase, peptide N-glycosidase F (PNGaseF), directly on followed by incubation releases amenable detection MALDI-IMS. The has been designed simultaneously released from intracellular organelle cell surface glycoproteins, while maintaining histopathology compatible...
A new mass spectrometry imaging approach to simultaneously map the two-dimensional distribution of N-glycans in tissues has been recently developed. The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) spatially profile location and multiple N-linked glycan species released by peptide N-glycosidase F frozen or formalin-fixed tissues. Multiple human hepatocellular carcinoma were evaluated with this method, resulting a panel over 30 detected. An...
Adeno-associated virus (AAV) vectors, which contain a DNA transgene packaged into protein capsid, have shown tremendous therapeutic potential in recent years. An inherent characteristic of the manufacturing process is production empty capsids that lack and are therefore unable to provide intended benefit. The effect on clinical outcomes not well understood, but there immunogenicity efficacy concerns, these considered product-related impurity. Therefore, should be controlled during monitored...
Abstract Glycosylation of cell surface, secreted, and circulating proteins is one the most common types post‐translational modification. These modifications occur commonly as three major classes: N‐linked glycosylation on asparagine residues, O‐linked serine or threonine glycosaminoglycan oligosaccharide polymers serine. Specifically, for glycans, an endoglycosidase enzyme, peptide N‐glycosidase F (PNGase F), cleaves attached oligosaccharides between first sugar. A method to analyze released...
The early detection of pancreatic ductal adenocarcinoma (PDAC) is a complex clinical obstacle yet key to improving the overall likelihood patient survival. Current and prospective carbohydrate biomarkers antigen 19-9 (CA19-9) sialylated tumor-related (sTRA) are sufficient for surveilling disease progression not approved delineating PDAC from other abdominal cancers noncancerous pathologies. To further understand these glycan epitopes, an imaging mass spectrometry (IMS) approach was used...
Recombinant adeno-associated virus (rAAV) has emerged as an important gene therapy vector with many clinical trials currently in progress. Analytical characterization and quantitation of particle content remain challenges both the development production rAAV vectors. In this study, charge detection mass spectrometry (CDMS) gel electrophoresis are used to characterize DNA recombinant AAV8 (rAAV8) vectors a wide range target genome sizes. We show that differences between masses empty particles...
Adeno-associated virus (AAV) is a widely used gene therapy vector. The intact packaged genome critical quality attribute and necessary for an effective therapeutic. In this work, charge detection mass spectrometry (CDMS) was to measure the molecular weight (MW) distribution of interest (GOI) extracted from recombinant AAV (rAAV) vectors. measured MWs were compared sequence masses range rAAV vectors with different GOIs, serotypes, production methods (Sf9 HEK293 cell lines). most cases,...
ABSTRACT A significant limitation of imaged capillary electric focusing (icIEF) is the inability to identify and characterize specific species in electropherogram. This has led development complementary ion‐exchange chromatography (IEX)‐based methods that are amenable either fraction collection subsequent characterization or online IEX coupled mass spectrometry. To overcome this while maintaining use icIEF, novel approaches, including an icIEF separation fractionation technology...
ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTHexahelicene. Absolute configurationDavid A. Lightner, Daniel T. Hefelfinger, Thomas W. Powers, Gerard Frank, and Kenneth N. TruebloodCite this: J. Am. Chem. Soc. 1972, 94, 10, 3492–3497Publication Date (Print):May 1, 1972Publication History Published online1 May 2002Published inissue 1 1972https://pubs.acs.org/doi/10.1021/ja00765a041https://doi.org/10.1021/ja00765a041research-articleACS PublicationsRequest reuse permissionsArticle...
Reducing the incidence and mortality rates for clear cell renal carcinoma (ccRCC) remains a significant clinical challenge with poor 5‐year survival rates. A unique tissue cohort was assembled of matched ccRCC distal nontumor tissues ( n = 20) associated moderate risk disease progression, half these from individuals who progressed to metastatic other remained free. These were used MALDI imaging MS profiling proteins in 2–20 kDa range, resulting panel 108 that had potential disease‐specific...
Using prostatic fluids rich in glycoproteins like prostate-specific antigen and acid phosphatase (PAP), the goal of this study was to identify structural types relative abundance glycans associated with prostate cancer status for subsequent use emerging MS-based glycopeptide analysis platforms.A series pooled samples expressed secretions (EPS) exosomes reflecting different stages disease were used N-linked glycan profiling by three complementary methods, MALDI-TOF profiling, normal-phase...
Adeno-associated viruses (AAVs) represent important gene therapy vectors with several approved clinical applications and numerous more in trials. Genome packaging is an essential step the bioprocessing of AAVs needs to be tightly monitored ensure proper delivery transgenes production effective drugs. Current methods monitor genome have limited sensitivity, a high demand on labor, struggle distinguish between intended or unwanted side-products. Here we show that Orbitrap-based...
Nearly one half of patients with lupus develop glomerulonephritis (GN), which often leads to renal failure. Although nephritis is diagnosed by the presence proteinuria, pathology can fall into five classes defined different forms tissue injury, and mechanisms involved in pathogenesis are not completely understood. Glycosphingolipids abundant kidney, have roles many cellular functions, were shown be other diseases. Here, we show dysfunctional glycosphingolipid metabolism MRL/lpr mice....
Clear-cell renal cell carcinoma (ccRCC) presents challenges to clinical management because of late-stage detection, treatment resistance, and frequent disease recurrence. Metabolically, ccRCC has a well-described Warburg effect utilization glucose, but how this affects complex carbohydrate synthesis alterations protein surface glycosylation is poorly defined. Using an imaging mass spectrometry approach, N-glycosylation patterns compositional differences were assessed between tumor nontumor...
The Multi-Attribute Method (MAM) Consortium was initially formed as a venue to harmonize best practices, share experiences, and generate innovative methodologies facilitate widespread integration of the MAM platform, which is an emerging ultra-high-performance liquid chromatography–mass spectrometry application. Successful implementation purity-indicating assay requires new peak detection (NPD) potential process- and/or product-related impurities. NPD interlaboratory study described herein...
Adeno-associated virus (AAV) gene therapy vectors, which contain a DNA transgene packaged into protein capsid, have shown tremendous therapeutic potential in recent years. Methods traditionally used quality control labs, such as high-performance liquid chromatography (HPLC) and capillary electrophoresis (CE), do not provide complete understanding of capsid viral (VP) charge heterogeneity. In the present study, we developed simple, one-step sample preparation charge-based VP separation using...
Discarded monkey autopsy specimens were used to investigate the mechanism of supracondylar hyperextension fracture. As fracture progressed from mild angulation complete lateral or medial displacement, anterior periosteum first was detached bone and then stripped distally before tearing over edge proximal fragment. Stability reduction by acute elbow flexion forearm pronation, owing compressive forces, transmitted through joint on side fracture, stability not significantly influenced a...
The ETS factor Friend leukemia virus integration 1 (FLI1) is a key modulator of lupus disease expression. Overexpressing FLI1 in healthy mice results the development an autoimmune kidney similar to that observed lupus. Lowering global levels two strains (Fli1(+/-)) significantly improved and prolonged survival. T cells from MRL/lpr Fli1(+/-) have reduced activation IL-4 production, neuraminidase expression, glycosphingolipid lactosylceramide. In this study, we demonstrate decreased renal...
The multi-attribute method (MAM), a recent advance in the application of liquid chromatography-mass spectrometry within pharmaceutical industry, enables simultaneous monitoring multiple product quality attributes single analytical method. While MAM is coupled with automated data processing and reporting, sample preparation, based on proteolytic peptide mapping, remains cumbersome low throughput. standard preparation for relies protein denaturation, reduction, alkylation prior to digestion,...