A5043022666- Extracellular vesicles in disease
- Viral Infectious Diseases and Gene Expression in Insects
- Monoclonal and Polyclonal Antibodies Research
- RNA Interference and Gene Delivery
- Biosimilars and Bioanalytical Methods
- CAR-T cell therapy research
- Virus-based gene therapy research
- CRISPR and Genetic Engineering
- Protein purification and stability
- MicroRNA in disease regulation
- Glycosylation and Glycoproteins Research
- Immunotherapy and Immune Responses
- interferon and immune responses
- Cell Adhesion Molecules Research
- Cancer Research and Treatments
- Cancer, Hypoxia, and Metabolism
- Nanoplatforms for cancer theranostics
- Biopolymer Synthesis and Applications
- Sexual Differentiation and Disorders
- Lysosomal Storage Disorders Research
- Radio Frequency Integrated Circuit Design
- Mass Spectrometry Techniques and Applications
- Cellular transport and secretion
- Galectins and Cancer Biology
- Autophagy in Disease and Therapy
Biogen (United States)
2013-2019
Protein Express (United States)
2007-2011
Massachusetts Institute of Technology
1997
Center for Cancer Research
1997
Roswell Park Comprehensive Cancer Center
1993-1995
University of Kentucky
1988
Extracellular vesicles (EVs) are an important intercellular communication system facilitating the transfer of macromolecules between cells. Delivery exogenous cargo tethered to EV surface or packaged inside lumen key strategies for generating therapeutic EVs. We identified two "scaffold" proteins, PTGFRN and BASP1, that preferentially sorted into EVs enable high-density display luminal loading a wide range molecules, including cytokines, antibody fragments, RNA binding vaccine antigens,...
Abstract Cyclic dinucleotide (CDN) agonists of the STimulator InterferoN Genes (STING) pathway have shown immune activation and tumor clearance in pre-clinical models. However, CDNs administered intratumorally also promote STING leading to direct cytotoxicity many cell types microenvironment (TME), systemic inflammation due rapid extravasation CDN, ablation TME. These result a failure establish immunological memory. ExoSTING, an engineered extracellular vesicle (EV) exogenously loaded with...
E2F directs the cell cycle-dependent expression of genes that induce or regulate division process. In mammalian cells, this transcriptional activity arises from combined properties multiple E2F-DP heterodimers. study, we show potential individual species is dependent upon their nuclear localization. This a constitutive property E2F-1, -2, and -3, whereas localization E2F-4 its association with other factors. We previously showed accounts for majority endogenous species. now subcellular...
Abstract Oncogenic osteomalacia (OOM) is associated with primitive mesenchymal tumors that secrete phosphaturic factors resulting in low serum concentrations of phosphate and calcitriol, phosphaturia, defective bone mineralization. To identify overexpressed genes these tumors, we compared gene expression profiles resected from patients OOM histologically similar control using serial analysis (SAGE). Three hundred sixty-four were expressed at least twofold greater tumors. A subset 67 highly...
Glycosylation in the Fc region of antibodies has been shown to play an important role antibody function. In current study, glycosylation human monoclonal was metabolically modulated using a potent alpha-mannosidase I inhibitor, kifunensine, resulting production with oligomannose-type N-glycans. Growing Chinese hamster ovary cells for 11 days batch culture single treatment kifunensine sufficient elicit this effect without any significant impact on cell viability or production. Antibodies...
The promise of IL12 as a cancer treatment has yet to be fulfilled with multiple tested approaches being limited by unwanted systemic exposure and unpredictable pharmacology. To address these limitations, we generated exoIL12, novel, engineered exosome therapeutic that displays functional on the surface an exosome. exosomal expression was achieved via fusion abundant protein PTGFRN resulting in equivalent potency vitro recombinant (rIL12) demonstrated IFNγ production. Following intratumoral...
One form of Niemann-Pick disease is caused by a deficiency in the enzymatic activity acid sphingomyelinase. During efforts to develop an enzyme replacement therapy based on recombinant human sphingomyelinase (rhASM), purified preparations were found have substantially increased specific if cell harvest media stored for several weeks at –20 °C prior purification. This increase was correlate with loss single free thiol rhASM, suggesting involvement cysteine residue. It demonstrated that...
Based on an optimized electroporation protocol, we designed a rapid, milliliter-scale diagnostic transient production assay to identify limitations in the ability of Chinese hamster ovary (CHO) cells produce model "difficult-to-express" homodimeric Fc-fusion protein, Sp35Fc, that exhibited very low volumetric titer and intracellular formation disulfide-bonded oligomeric aggregates post-transfection. As expression Sp35Fc induced unfolded protein response transfected host cells, utilized...
Flow cytometry was partnered with a nonfluorescent reporter protein for rapid, early stage identification of clones producing high levels therapeutic protein. A cell surface protein, not normally expressed on CHO cells, is coexpressed, as reporter, the and detected using fluorescently labeled antibody. The genes encoding are linked by an IRES, so that they transcribed in same mRNA but translated independently. Since each arise from common mRNA, protein's expression level accurately predicts...
The Biogen upstream platform is capable of delivering equivalent quality material throughout the cell line generation process. This allows us to rapidly deliver high-quality biopharmaceuticals patients with unmet medical needs. drive reduce time-to-market led engineering group develop an expression system that can enable this strategy. We have developed a clonal Chinese Hamster Ovary (CHO) host routinely produce consistent antibody at high titers enables faster delivery early phase through...
A central goal for most biopharmaceutical companies is to reduce the development timeline reach clinical proof of concept. This objective requires tools that ensure quality biotherapeutic material destined clinic. Recent advances in high throughput protein analytics provide confidence our ability assess productivity and product attributes at early stages cell line development. However, one attribute has, until recently, been absent from standard battery analytical tests facilitating informed...
This case study addresses the difficulty in achieving high level expression and production of a small, very positively charged recombinant protein. The novel challenges with this protein include protein's adherence to cell surface its inhibitory effects on Chinese hamster ovary (CHO) growth. To overcome these challenges, we utilized multi-prong approach. We identified dextran sulfate as way simultaneously extract from boost cellular productivity. In addition, host cells were adapted grow...
Cells exposed to hypoxia undergo substantial changes in gene expression generally associated with metabolic adaptation and increasing oxygen delivery. In contrast, responses distinct from those elicited by are induced anoxic fibroblasts; this includes activation of a set VL30 elements. The seen anoxically cultured fibroblasts expressed physiologically vivo during the anaerobic phase wound healing. A fundamental question is whether transcriptional regulatory pathways utilized anoxia already...
Here we describe a method that couples flow cytometric detection with the attenuated translation of reporter protein to enable efficient selection CHO clones producing high levels recombinant proteins. In this system, small cell surface is expressed from an upstream open reading frame utilizing non-AUG initiation (alternate start) codon. Due low efficiency alternate start codon, majority events occur at first AUG downstream encoding interest. While significantly reduced, are sufficient for...
We developed a novel reversed-phase liquid chromatography method for monitoring multiple ingredients in poloxamers and evaluating their stability.
Abstract Background: The Stimulator of Interferon Genes (STING) pathway is an attractive target in immuno-oncology. Selective activation the STING antigen presenting cells (APCs) essential for eliciting a potent and specific anti-tumor immune response. ubiquitously expressed normal including T endothelial cells. Direct intra-tumoral administration free agonist results all cells, resulting loss viability tissue damage, systemic due to vascular leakage. Exosomes are efficient natural messenger...
The serum half-life, biological activity, and solubility of many recombinant glycoproteins depend on their sialylation. Monitoring glycoprotein sialylation during cell culture manufacturing is, therefore, critical to ensure product efficacy safety. Here a high-throughput method for semi-quantitative fingerprinting using capillary isoelectric focusing immunoassay NanoPro (Protein Simple) platform was developed. specific, sensitive, precise, robust. It could analyze 2 μL crude samples without...
VL30 elements are associated with cancer by their overexpression in rodent malignancies, induction a fibroblast response to anoxia which shares features the malignant phenotype, and presence recombined into Harvey murine sarcoma virus (HaSV) Kirsten virus. These viruses contain ras oncogenes flanked on both sides retrotransposon element sequences, turn mouse leukemia sequences. Three very basic questions have existed about sequences found viruses: (i) how did they become recombined, (ii)...