keyword
MENU ▼
Read by QxMD icon Read
search

biotherapeutic

keyword
https://www.readbyqxmd.com/read/28342833/a-high-density-cho-s-transient-transfection-system-comparison-of-expicho-and-expi293
#1
Nina K Jain, Susan Barkowski-Clark, Richard Altman, Krista Johnson, Fang Sun, Jonathan Zmuda, Chao Yan Liu, Adriana Kita, Ryan Schulz, Alyssa Neill, Robert Ballinger, Rekha Patel, Jian Liu, Alinafe Mpanda, Brian Huta, Henry Chiou, Walter Voegtli, Tadas Panavas
Chinese Hamster Ovary (CHO) cells are the principal mammalian host used for stable cell line generation and biotherapeutic protein production. Until recently, production of milligrams to grams of protein in CHO transient systems was challenging. As such, Human Embryonic Kidney (HEK293) cells are the most common mammalian cell type used for transient transfection. The post-translational modifications (PTMs) of a protein are dictated in part by the cell line used for expression, and changes in PTMs have been shown to affect both the activity and biophysical properties of proteins...
March 22, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28328036/use-of-a-stable-isotope-labeled-reporter-peptide-and-antioxidants-for-reliable-quantification-of-methionine-oxidation-in-a-monoclonal-antibody-by-liquid-chromatography-mass-spectrometry
#2
Pilsoo Kang, Tanya Mezhebovsky, Wei Chen, James A McCardle, Sheng Zhang, Eric P Sullivan, Philip M Sass, Eric Routhier
RATIONALE: Accurate quantification of methionine oxidation in therapeutic proteins by liquid chromatography/mass spectrometry (LC/MS) is challenging due to the potential artifacts introduced during sample preparation and analysis in the peptide mapping workflow. In this study, a systematic approach for optimization of the peptide mapping procedure to achieve reliable quantification of endogenous methionine oxidation in monoclonal antibodies was developed. METHODS: The approach is based on usage of a stable-isotope-labeled reporter peptide, identical in sequence to the tryptic peptide of an IgG1 monoclonal antibody containing the methionine residue most prone to oxidation...
July 30, 2016: Rapid Communications in Mass Spectrometry: RCM
https://www.readbyqxmd.com/read/28289128/characterizing-the-overall-derivatization-of-conjugated-oligomeric-proteins
#3
Michaela Wendeler, Wei Zhao, Christopher Thompson, Manuel Baca, Dhanesh Gadre, Xiangyang Wang, Harry Yang
The analysis and accurate quantitation of bioconjugations proves challenging in the case of oligomeric proteins, especially when the size of the molecule or the nature of the conjugate do not allow the analysis of the intact protein under native conditions. In this case, analytical methods are frequently applied that result in a dissociation of non-covalently linked subunits. This limits the analysis to a description of individual subunits, thereby obscuring the accurate characterization of the overall functionalization...
March 13, 2017: PDA Journal of Pharmaceutical Science and Technology
https://www.readbyqxmd.com/read/28267992/clostridium-difficile-disease-diagnosis-pathogenesis-and-treatment-update
#4
REVIEW
Lena M Napolitano, Charles E Edmiston
Clostridium difficile infections are the leading cause of health care-associated infectious diarrhea, posing a significant risk for both medical and surgical patients. Because of the significant morbidity and mortality associated with C difficile infections, knowledge of the epidemiology of C difficile in combination with a high index of suspicion and susceptible patient populations (including surgical, postcolectomy, and inflammatory bowel disease patients) is warranted. C difficile infections present with a wide spectrum of disease, ranging from mild diarrhea to fulminant colitis or small bowel enteritis and recurrent C difficile infections...
March 3, 2017: Surgery
https://www.readbyqxmd.com/read/28255747/mapping-the-binding-interface-of-vegf-and-a-monoclonal-antibody-fab-1-fragment-with-fast-photochemical-oxidation-of-proteins-fpop-and-mass-spectrometry
#5
Ying Zhang, Aaron T Wecksler, Patricia Molina, Galahad Deperalta, Michael L Gross
We previously analyzed the Fab-1:VEGF (vascular endothelial growth factor) system described in this work, with both native top-down mass spectrometry and bottom-up mass spectrometry (carboxyl-group or GEE footprinting) techniques. This work continues bottom-up mass spectrometry analysis using a fast photochemical oxidation of proteins (FPOP) platform to map the solution binding interface of VEGF and a fragment antigen binding region of an antibody (Fab-1). In this study, we use FPOP to compare the changes in solvent accessibility by quantitating the extent of oxidative modification in the unbound versus bound states...
March 2, 2017: Journal of the American Society for Mass Spectrometry
https://www.readbyqxmd.com/read/28249362/effects-of-lysosomal-biotherapeutic-recombinant-protein-expression-on-cell-stress-and-protease-and-general-host-cell-protein-hcp-release-in-chinese-hamster-ovary-cells
#6
Damiano Migani, C Mark Smales, Daniel G Bracewell
Recombinant human Acid Alpha Glucosidase (GAA) is the therapeutic enzyme used for the treatment of Pompe disease, a rare genetic disorder characterised by GAA deficiency in the cell lysosomes(1) . The manufacturing process for GAA can be challenging, in part due to protease degradation. The overall goal of this study was to understand the effects of GAA overexpression on cell lysosomal phenotype and host cell protein (HCP) release, and any resultant consequences for protease levels and ease of manufacture. To do this we first generated a human recombinant GAA producing stable CHO cell line and designed the capture chromatographic step anion exchange (IEX)...
March 1, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28242452/top-down-analysis-of-immunoglobulin-g-isotypes-1-and-2-with-electron-transfer-dissociation-on-a-high-field-orbitrap-mass-spectrometer
#7
Luca Fornelli, Daniel Ayoub, Konstantin Aizikov, Xiaowen Liu, Eugen Damoc, Pavel A Pevzner, Alexander Makarov, Alain Beck, Yury O Tsybin
The increasing importance of immunoglobulins G (IgGs) as biotherapeutics calls for improved structural characterization methods designed for these large (~150kDa) macromolecules. Analysis workflows have to be rapid, robust, and require minimal sample preparation. In a previous work we showed the potential of Orbitrap Fourier transform mass spectrometry (FTMS) combined with electron transfer dissociation (ETD) for the top-down investigation of an intact IgG1, resulting in ~30% sequence coverage. Here, we describe a top-down analysis of two IgGs1 (adalimumab and trastuzumab) and one IgG2 (panitumumab) performed with ETD on a mass spectrometer equipped with a high-field Orbitrap mass analyzer...
February 24, 2017: Journal of Proteomics
https://www.readbyqxmd.com/read/28240851/ion-mobility-mass-spectrometry-reveals-a-dipeptide-that-acts-as-a-molecular-chaperone-for-amyloid-%C3%AE
#8
Molly T Soper-Hopper, Joseph D Eschweiler, Brandon T Ruotolo
Previously, we discovered and structurally characterized a complex between amyloid β 1-40 and the neuropeptide leucine enkephalin. This work identified leucine enkephalin as a potentially useful starting point for the discovery of peptide-related biotherapeutics for Alzheimer's disease. In order to better understand such complexes that are formed in vitro, we describe here the analysis of a series of site-directed amino acid substitution variants of both peptides, covering the leucine enkephalin sequence in its entirety and a large number of selected residues of amyloid β 1-40 (residues: D1, E3, F4, R5, H6, Y10, E11, H13, H14, Q15, K16, E22, K28, and V40)...
February 27, 2017: ACS Chemical Biology
https://www.readbyqxmd.com/read/28237613/effects-of-probiotics-supplement-in-patients-with-type-2-diabetes-mellitus-a-meta-analysis-of-randomized-trials
#9
REVIEW
Yi-Meng Hu, Feng Zhou, Yin Yuan, Yan-Cheng Xu
BACKGROUND AND OBJECTIVE: To objectively evaluate the effects of probiotics supplement on glycemic control and lipid metabolism in patients with type 2 diabetes mellitus (T2DM). MATERIAL AND METHODS: The randomized controlled trials (RCTs) with regard to the probiotics or synbiotics for the treatment of T2DM were collected through retrieving 5 databases from their establishment to March 2016. After study selection, quality assessment and data extraction were performed by 2 authors independently; and STATA software was used for statistical analysis...
February 22, 2017: Medicina Clínica
https://www.readbyqxmd.com/read/28223164/exploring-sensitivity-throughput-of-a-parallel-flow-spri-biosensor-for-characterization-of-antibody-antigen-interaction
#10
Vishal Kamat, Ashique Rafique
Rapid growth in the field of biotherapeutics has led to an increased demand for high-throughput, label-free biosensors exhibiting high sensitivity. To support the current needs, Sierra Sensors introduced a surface plasmon resonance imaging (SPRi) based biosensor, Molecular Affinity Screening System (MASS-1). We assessed the potential utility of MASS-1 to support Regeneron's therapeutic antibody discovery. A large panel of antibody-antigen interactions was characterized using MASS-1 and the kinetic data were compared with the Biacore 4000 biosensor...
February 20, 2017: Analytical Biochemistry
https://www.readbyqxmd.com/read/28212986/correlating-the-effects-of-antimicrobial-preservatives-on-conformational-stability-aggregation-propensity-and-backbone-flexibility-of-an-igg1-mab
#11
Jayant Arora, Sangeeta B Joshi, C Russell Middaugh, David D Weis, David B Volkin
Multidose formulations of biotherapeutics, which offer better dosage management and reduced production costs, require the addition of antimicrobial preservatives (APs). APs have been shown, however, to decrease protein stability in solution and cause protein aggregation. In this report, the effect of 4 APs, m-cresol, phenol, phenoxyethanol, and benzyl alcohol on conformational stability, aggregation propensity, and backbone flexibility of an IgG1 mAb, mAb-4, is investigated. Compared with no preservative control, each of the APs decreased the conformational stability of mAb-4 as measured by differential scanning calorimetry and extrinsic fluorescence spectroscopy...
February 14, 2017: Journal of Pharmaceutical Sciences
https://www.readbyqxmd.com/read/28210903/role-of-exchange-protein-directly-activated-by-camp-epac1-in-breast-cancer-cell-migration-and-apoptosis
#12
Naveen Kumar, Sonal Gupta, Surbhi Dabral, Shailja Singh, Seema Sehrawat
Despite the current progress in cancer research and therapy, breast cancer remains the leading cause of mortality among half a million women worldwide. Migration and invasion of cancer cells are associated with prevalent tumor metastasis as well as high mortality. Extensive studies have powerfully established the role of prototypic second messenger cAMP and its two ubiquitously expressed intracellular cAMP receptors namely the classic protein kinaseA/cAMP-dependent protein kinase (PKA) and the more recently discovered exchange protein directly activated by cAMP/cAMP-regulated guanine nucleotide exchange factor (EPAC/cAMP-GEF) in cell migration, cell cycle regulation, and cell death...
February 16, 2017: Molecular and Cellular Biochemistry
https://www.readbyqxmd.com/read/28197641/m%C3%A3-llerian-inhibiting-substance-inhibits-an-ovarian-cancer-cell-line-via-%C3%AE-catenin-interacting-protein-deregulation-of-the-wnt-signal-pathway
#13
Sang Ho Park, Youn Jee Chung, Jae Yen Song, Sang Il Kim, David Pépin, David T MacLaughlin, Patricia K Donahoe, Jang Heub Kim
Müllerian inhibiting substance/anti-Müllerian hormone (MIS/AMH) has been suggested as a biotherapeutic agent in gynecological cancers that highly express the MIS/AMH type II receptors (MISRII/AMHRII) but the anticancer mechanisms by which MIS/AMH acts are not fully understood. Our experiments show that MIS/AMH inhibits ovarian cancer by deregulating the Wnt signal pathway via the β-catenin interacting protein (ICAT). MIS/AMH inhibition of ICAT by small interfering RNAs (siRNA) decreased ICAT driven ovarian cancer cell viability as measured by the methylthiazoltetrazolium assay, reversed cell cycle arrest and annexin V expression and diminished migration by scratch wound assay...
March 2017: International Journal of Oncology
https://www.readbyqxmd.com/read/28192936/generic-hybrid-ligand-binding-assay-liquid-chromatography-high-resolution-mass-spectrometry-based-workflow-for-multiplexed-human-immunoglobulin-g1-quantification-at-the-intact-protein-level-application-to-preclinical-pharmacokinetic-studies
#14
Christian Lanshoeft, Sarah Cianférani, Olivier Heudi
The quantitative analysis of human immunoglobulin G1 (hIgG1) by mass spectrometry is commonly performed using surrogate peptides after enzymatic digestion. Since some limitations are associated with this approach, a novel workflow is presented by hybridizing ligand binding assay (LBA) with liquid chromatography-high-resolution mass spectrometry (LC-HRMS) for hIgG1 quantification directly at the intact protein level. Different hIgG1s, including a [(13)C]-labeled version used as internal standard, were immuno-enriched from rat serum with a fully automated platform based on streptavidin coated tips and a biotinylated mouse anti-hIgG capture antibody targeting the fragment crystallizable region followed by overnight deglycosylation prior to LC-HRMS analysis...
February 9, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28192904/high-resolution-glycan-analysis-by-temperature-gradient-capillary-electrophoresis
#15
Marton Szigeti, Andras Guttman
Temperature gradient capillary electrophoresis was introduced to enhance separation selectivities for branched glycans of biotherapeutic interest. A mixture of afucosylated, fucosylated, and high mannose oligosaccharides was separated in the range of 15 to 45 °C at 5 °C temperature intervals. It was found that within this temperature range, the separation selectivity was carbohydrate structure dependent. The resolution between some glycan structures was greater at elevated temperatures, while others separated better at lower temperatures...
February 3, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28186334/a-novel-bxb1-integrase-rmce-system-for-high-fidelity-site-specific-integration-of-mab-expression-cassette-in-cho-cells
#16
Mara C Inniss, Kalpanie Bandara, Barbara Jusiak, Tim K Lu, Ron Weiss, Liliana Wroblewska, Lin Zhang
As CHO cell line development for biotherapeutic production becomes more sophisticated through the availability of the CHO genome sequence, the ability to accurately and reproducibly engineer the host cell genome has become increasingly important. Multiple well characterized systems for site-specific integration will enable more complex cell line engineering to generate cell lines with desirable attributes. We built and characterized a novel recombinase mediated cassette exchange (RMCE) system using Bxb1 integrase and compared it to the commonly used Flp/FRT RMCE system...
February 10, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28174761/detection-of-glycosylation-and-iron-binding-protein-modifications-using-raman-spectroscopy
#17
Lorna Ashton, Victoria L Brewster, Elon Correa, Royston Goodacre
In this study we demonstrate the use of Raman spectroscopy to determine protein modifications as a result of glycosylation and iron binding. Most proteins undergo some modifications after translation which can directly affect protein function. Identifying these modifications is particularly important in the production of biotherapeutic agents as they can affect stability, immunogenicity and pharmacokinetics. However, post-translational modifications can often be difficult to detect with regard to the subtle structural changes they induce in proteins...
February 8, 2017: Analyst
https://www.readbyqxmd.com/read/28160363/establishment-of-a-tobacco-by2-cell-line-devoid-of-plant-specific-xylose-and-fucose-as-a-platform-for-the-production-of-biotherapeutic-proteins
#18
Uri Hanania, Tami Ariel, Yoram Tekoah, Liat Fux, Maor Sheva, Yehuda Gubbay, Mara Weiss, Dina Oz, Yaniv Azulay, Albina Turbovski, Yehava Forster, Yoseph Shaaltiel
Plant produced glycoproteins contain N-linked glycans with plant-specific residues of β(1,2)-xylose and core α(1,3)-fucose, which do not exist in mammalian derived proteins. Although our experience with two enzymes that are used for enzyme replacement therapy (ERT) does not indicate that the plant sugar residues have deleterious effects, we made a conscious decision to eliminate these moieties from plant-expressed proteins. We knocked out the β(1,2)-xylosyltranferase (XylT) and the α(1,3)-fucosyltransferase (FucT) genes, using CRISPR/Cas9 genome editing, in Nicotiana...
February 3, 2017: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/28150882/detection-and-measurement-of-methionine-oxidation-in-proteins
#19
K Ilker Sen, Robert Hepler, Hirsh Nanda
Methionine oxidation is a prevalent modification found in proteins both in biological settings and in the manufacturing of biotherapeutic molecules. In cells, the oxidation of specific methionine sites can modulate protein function or promote interactions that trigger signaling pathways. In biotherapeutic development, the formation of oxidative species could be detrimental to the efficacy or safety of the drug product. Thus, methionine oxidation is a critical quality attribute that needs to be monitored throughout development...
February 2, 2017: Current Protocols in Protein Science
https://www.readbyqxmd.com/read/28138368/investigating-structure-and-dynamics-of-proteins-in-amorphous-phases-using-neutron-scattering
#20
REVIEW
Maria Monica Castellanos, Arnold McAuley, Joseph E Curtis
In order to increase shelf life and minimize aggregation during storage, many biotherapeutic drugs are formulated and stored as either frozen solutions or lyophilized powders. However, characterizing amorphous solids can be challenging with the commonly available set of biophysical measurements used for proteins in liquid solutions. Therefore, some questions remain regarding the structure of the active pharmaceutical ingredient during freezing and drying of the drug product and the molecular role of excipients...
2017: Computational and Structural Biotechnology Journal
keyword
keyword
106671
1
2
Fetch more papers »
Fetching more papers... Fetching...
Read by QxMD. Sign in or create an account to discover new knowledge that matter to you.
Remove bar
Read by QxMD icon Read
×

Search Tips

Use Boolean operators: AND/OR

diabetic AND foot
diabetes OR diabetic

Exclude a word using the 'minus' sign

Virchow -triad

Use Parentheses

water AND (cup OR glass)

Add an asterisk (*) at end of a word to include word stems

Neuro* will search for Neurology, Neuroscientist, Neurological, and so on

Use quotes to search for an exact phrase

"primary prevention of cancer"
(heart or cardiac or cardio*) AND arrest -"American Heart Association"