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https://www.readbyqxmd.com/read/27914608/evaluation-of-size-exclusion-chromatography-columns-packed-with-sub-3%C3%AE-m-particles-for-the-analysis-of-biopharmaceutical-proteins
#1
Alexandre Goyon, Alain Beck, Olivier Colas, Koen Sandra, Davy Guillarme, Szabolcs Fekete
The aim of this study was to evaluate the practical possibilities and limitations of several recently introduced size exclusion chromatographic (SEC) columns of 150×4.6mm, sub-3μm (Agilent AdvanceBioSEC 2.7μm, Tosoh TSKgel UP-SW3000 2.0μm, Phenomenex Yarra SEC X-150 1.8μm and Waters Acquity BEH200 1.7μm) for the separation of biopharmaceutical proteins. For this purpose, some model proteins were tested, as well as several commercial therapeutic monoclonal antibodies (mAbs) and antibody-drug-conjugates (ADCs)...
November 27, 2016: Journal of Chromatography. A
https://www.readbyqxmd.com/read/27855600/optimization-on-fc-for-improvement-of-stability-and-aggregation-resistance
#2
Xiaobo Chen, Fang Zeng, Tao Huang, Liang Cheng, Huan Liu, Rui Gong
Fc-based therapeutics including therapeutic full-size monoclonal antibodies (mAbs) and Fc-fusion proteins represent fastest-growing market in biopharmaceutical industrial. However, one major challenge during development of Fc-based therapeutics is how to maintain their efficacy in clinic use. Many factors may lead to failure in final marketing. For example, the stability and aggregation resistance might not be high enough for bearing the disadvantages during fermentation, purification, formulation, storage, shipment and other steps in manufacture and sale...
November 17, 2016: Current Pharmaceutical Biotechnology
https://www.readbyqxmd.com/read/27789805/processing-impact-on-monoclonal-antibody-drug-products-protein-subvisible-particulate-formation-induced-by-grinding-stress
#3
Benson Gikanga, Devon Roshan-Eisner, Robert Ovadia, Eric S Day, Oliver Boris Stauch, Yuh-Fun Maa
Subvisible particle formation in monoclonal antibody (mAb) drug product resulting from mixing and filling operations represents a significant processing risk that can lead to filter fouling and thereby lead to process delays or failures. Several previous studies from our lab and others demonstrated the formation of subvisible particulates in mAb formulations resulting from mixing operations using some bottom-mounted mixers or stirrer bars. It was hypothesized that the stress (e.g. shear/cavitation) derived from tight clearance and/or close contact between the impeller and shaft was responsible for SvP generation...
October 27, 2016: PDA Journal of Pharmaceutical Science and Technology
https://www.readbyqxmd.com/read/27788030/pmab-38-recognizes-canine-podoplanin-of-squamous-cell-carcinomas
#4
Mika K Kaneko, Ryusuke Honma, Satoshi Ogasawara, Yuki Fujii, Takuro Nakamura, Noriko Saidoh, Michiaki Takagi, Yumiko Kagawa, Satoru Konnai, Yukinari Kato
Podoplanin, a type I transmembrane protein, is expressed in lymphatic endothelial cells. Although we previously developed an anticanine podoplanin monoclonal antibody (mAb), PMab-38, immunohistochemistry (IHC) showed that it did not react with canine lymphatic endothelial cells. Here, we determined whether PMab-38 recognizes canine podoplanin of squamous cell carcinomas (SCCs) and clarified its epitope. In IHC, PMab-38 reacted with 83% of SCCs (15/18 cases). Flow cytometry showed that the epitope of PMab-38 was different from that of the platelet aggregation-stimulating domain of the N-terminus, which was detected by almost all antipodoplanin mAbs such as D2-40 or NZ-1...
October 2016: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/27699633/antibody-based-drugs-and-approaches-against-amyloid-%C3%AE-species-for-alzheimer-s-disease-immunotherapy
#5
Jing Liu, Bin Yang, Jun Ke, Wenjia Li, Wen-Chen Suen
Alzheimer's disease (AD), one of the most devastating diseases for the older population, has become a major healthcare burden in the increasingly aging society worldwide. Currently, there are still only symptomatic treatments available on the market, just to slow down disease progression. In the past decades, extensive research focusing on the development of immunotherapy using monoclonal antibodies (mAbs) as potential "disease-modifying drugs" has shown promise in inhibiting or clearing the formation of toxic amyloid-β (Aβ) species, the suspected causative agents of AD...
October 2016: Drugs & Aging
https://www.readbyqxmd.com/read/27697310/comprehensive-study-on-the-effects-of-sodium-and-potassium-additives-in-size-exclusion-chromatographic-separations-of-protein-biopharmaceuticals
#6
Alexandre Goyon, Alain Beck, Jean-Luc Veuthey, Davy Guillarme, Szabolcs Fekete
To separate proteins solely based on their difference in hydrodynamic volume in size exclusion chromatography (SEC), the ionic strength of the mobile phase has to be increased in order to avoid secondary ionic interactions between proteins and the stationary phase. However, adding salts to the mobile phase can have a serious effect on protein aggregation and can lead to artifacts. In the present study, several monoclonal antibodies (mAbs) and the antibody-drug conjugate (ADC), trastuzumab emtansine were selected to study the effect of mobile phase salt additive on aggregation measurements...
September 26, 2016: Journal of Pharmaceutical and Biomedical Analysis
https://www.readbyqxmd.com/read/27660109/choosing-the-right-protein-a-affinity-chromatography-media-can-remove-aggregates-efficiently
#7
Tomokazu Yada, Koichi Nonaka, Masayuki Yabuta, Noriko Yoshimoto, Shuichi Yamamoto
Protein A chromatography (PAC) is commonly used as an efficient capture step in monoclonal antibody (mAb) separation processes. Usually dynamic binding capacity is used for choosing the right PAC. However, if aggregates can be efficiently removed during elution, it can make the following polishing steps easier. In this study a method for choosing the right PAC media in terms of mAb aggregate removal is proposed. Linear pH gradient elution experiments of two different mAbs on various PAC columns are carried out, where the elution behavior of aggregates as well as the monomer is measured...
September 23, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27643824/a-method-to-measure-protein-unfolding-at-an-air-liquid-interface
#8
Danielle L Leiske, Ian C Shieh, Martha Lovato Tse
Proteins are surface-active molecules that have a propensity to adsorb to hydrophobic interfaces, such as the air-liquid interface. Surface flow can increase aggregation of adsorbed proteins, which may be an undesirable consequence depending on the application. As changes in protein conformation upon adsorption are thought to induce aggregation, the ability to measure the folded state of proteins at interfaces is of particular interest. However, few techniques currently exist to measure protein conformation at interfaces...
October 4, 2016: Langmuir: the ACS Journal of Surfaces and Colloids
https://www.readbyqxmd.com/read/27589351/characterizing-monoclonal-antibody-formulations-in-arginine-glutamate-solutions-using-1-h-nmr-spectroscopy
#9
Priscilla Kheddo, Matthew J Cliff, Shahid Uddin, Christopher F van der Walle, Alexander P Golovanov
Assessing how excipients affect the self-association of monoclonal antibodies (mAbs) requires informative and direct in situ measurements for highly concentrated solutions, without sample dilution or perturbation. This study explores the application of solution nuclear magnetic resonance (NMR) spectroscopy for characterization of typical mAb behavior in formulations containing arginine glutamate. The data show that the analysis of signal intensities in 1D (1)H NMR spectra, when compensated for changes in buffer viscosity, is invaluable for identifying conditions where protein-protein interactions are minimized...
August 11, 2016: MAbs
https://www.readbyqxmd.com/read/27568167/a-simple-and-efficient-purification-platform-for-monoclonal-antibody-production-based-on-chromatin-directed-cell-culture-clarification-integrated-with-precipitation-and-void-exclusion-anion-exchange-chromatography
#10
Quan Chen, Sarah Maria Abdul Latiff, Phyllicia Toh, Xinying Peng, Aina Hoi, Mo Xian, Haibo Zhang, Rui Nian, Wei Zhang, Pete Gagnon
Protein A affinity chromatography, featured by its robustness and high-specificity, is still dominant as a first capture step for the purification of immunoglobulin G monoclonal antibodies (IgG mAbs). However, the material and operational costs of protein A are universally recognized as high, and its productivity is also limited as column mode. In order to overcome these limitations, industry is increasingly considering the use of non-protein A-based processes for IgG purification. In this study, sodium citrate precipitation (SCP) was developed as the primary purification step, and chromatin-directed cell culture clarification was demonstrated to significantly elevate the purification capability...
October 20, 2016: Journal of Biotechnology
https://www.readbyqxmd.com/read/27564251/lpmab-19-recognizes-sialylated-o-glycan-on-thr76-of-human-podoplanin
#11
Satoshi Ogasawara, Mika K Kaneko, Yukinari Kato
Human podoplanin (hPDPN) is expressed in lymphatic vessels, pulmonary type-I alveolar cells, and renal glomerulus. The hPDPN/C-type lectin-like receptor-2 (CLEC-2) interaction is involved in platelet aggregation and cancer metastasis. High expression of hPDPN in cancer cells or cancer-associated fibroblasts (CAFs) leads to a poor prognosis for cancer patients. In our previous research, we reported on several anti-hPDPN monoclonal antibodies (mAbs), including LpMab-2, LpMab-3, LpMab-7, LpMab-9, LpMab-12, LpMab-13, and LpMab-17 of mouse IgG1 subclass, which were produced using CasMab technology...
August 26, 2016: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/27494246/use-of-in-vitro-assays-to-assess-immunogenicity-risk-of-antibody-based-biotherapeutics
#12
Marisa K Joubert, Meghana Deshpande, Jane Yang, Helen Reynolds, Christine Bryson, Mark Fogg, Matthew P Baker, Jonathan Herskovitz, Theresa J Goletz, Lei Zhou, Michael Moxness, Gregory C Flynn, Linda O Narhi, Vibha Jawa
An In Vitro Comparative Immunogenicity Assessment (IVCIA) assay was evaluated as a tool for predicting the potential relative immunogenicity of biotherapeutic attributes. Peripheral blood mononuclear cells from up to 50 healthy naïve human donors were monitored up to 8 days for T-cell proliferation, the number of IL-2 or IFN-γ secreting cells, and the concentration of a panel of secreted cytokines. The response in the assay to 10 monoclonal antibodies was found to be in agreement with the clinical immunogenicity, suggesting that the assay might be applied to immunogenicity risk assessment of antibody biotherapeutic attributes...
2016: PloS One
https://www.readbyqxmd.com/read/27484800/ligand-induced-epitope-masking-dissociation-of-integrin-%C3%AE-5%C3%AE-1-fibronectin-complexes-only-by-monoclonal-antibodies-with-an-allosteric-mode-of-action
#13
A Paul Mould, Janet A Askari, Adam Byron, Yoshikazu Takada, Thomas A Jowitt, Martin J Humphries
We previously demonstrated that Arg-Gly-Asp (RGD)-containing ligand-mimetic inhibitors of integrins are unable to dissociate pre-formed integrin-fibronectin complexes (IFCs). These observations suggested that amino acid residues involved in integrin-fibronectin binding become obscured in the ligand-occupied state. Because the epitopes of some function-blocking anti-integrin monoclonal antibodies (mAbs) lie near the ligand-binding pocket, it follows that the epitopes of these mAbs may become shielded in the ligand-occupied state...
September 30, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27452807/effect-of-charge-localization-on-the-in-vivo-optical-imaging-properties-of-near-infrared-cyanine-dye-monoclonal-antibody-conjugates
#14
Kazuhide Sato, Alexander P Gorka, Tadanobu Nagaya, Megan S Michie, Yuko Nakamura, Roger R Nani, Vince L Coble, Olga V Vasalatiy, Rolf E Swenson, Peter L Choyke, Martin J Schnermann, Hisataka Kobayashi
Near-infrared (NIR) fluorophores show superior in vivo imaging properties than visible-light fluorophores because of the increased light penetration in tissue and lower autofluorescence of these wavelengths. We have recently reported that new NIR cyanine dyes containing a novel C4'-O-alkyl linker exhibit greater chemical stability and excellent optical properties relative to existing C4'-O-aryl variants. In this study, we synthesized two NIR cyanine dyes with the same core structure and charge but different indolenine substituents: FNIR-Z-759 bearing a combination of two sulfonates and two quaternary ammonium cations, and FNIR-G-765 bearing a combination of two sulfonates and two guanidines, resulting in zwitterionic charge with distinct cationic moieties...
October 20, 2016: Molecular BioSystems
https://www.readbyqxmd.com/read/27385782/dimeric-fc%C3%AE-r-ectodomains-as-probes-of-the-fc-receptor-function-of-anti-influenza-virus-igg
#15
Bruce D Wines, Hillary A Vanderven, Sandra E Esparon, Anne B Kristensen, Stephen J Kent, P Mark Hogarth
Ab-dependent cellular cytotoxicity, phagocytosis, and Ag presentation are key mechanisms of action of Abs arising in vaccine or naturally acquired immunity, as well of therapeutic mAbs. Cells expressing the low-affinity FcγRs (FcγRII or CD32 and FcγRIII or CD16) are activated for these functions when receptors are aggregated following the binding of IgG-opsonized targets. Despite the diversity of the Fc receptor proteins, IgG ligands, and potential responding cell types, the induction of all FcγR-mediated responses by opsonized targets requires the presentation of multiple Fc regions in close proximity to each other...
August 15, 2016: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/27380437/effects-of-protein-conformation-apparent-solubility-and-protein-protein-interactions-on-the-rates-and-mechanisms-of-aggregation-for-an-igg1monoclonal-antibody
#16
Cavan Kalonia, Vishal Toprani, Ronald Toth, Newton Wahome, Ian Gabel, C Russell Middaugh, David B Volkin
Non-native protein aggregation is a key degradation pathway of immunoglobulins. In this work, the aggregation kinetics of an immunoglobulin gamma-1 monoclonal antibody (IgG1 mAb) in different solution environments was monitored over a range of incubation temperatures for up to seven months using size exclusion chromatography. Histidine and citrate buffers with/without sodium chloride were employed to modulate the mAb's conformational stability, solubility (in the presence of polyethylene glycol, PEG), and protein-protein interactions as measured by differential scanning calorimetry, PEG precipitation, and static light scattering, respectively...
July 28, 2016: Journal of Physical Chemistry. B
https://www.readbyqxmd.com/read/27380308/specific-detection-of-dog-podoplanin-expressed-in-renal-glomerulus-by-a-novel-monoclonal-antibody-pmab-38-in-immunohistochemistry
#17
Ryusuke Honma, Mika K Kaneko, Satoshi Ogasawara, Yuki Fujii, Satoru Konnai, Michiaki Takagi, Yukinari Kato
Podoplanin (PDPN) is expressed in several normal tissues including podocytes of renal glomerulus, lymphatic endothelial cells (LECs), and type I alveolar cells of lung. PDPN activates platelet aggregation by binding to C-type lectin-like receptor-2 (CLEC-2) on platelets. Many monoclonal antibodies (mAbs) against human PDPN, mouse PDPN, rat PDPN, rabbit PDPN, and bovine PDPN have been established; antidog PDPN (dPDPN) mAbs have not been developed. Herein, we immunized mice with the recombinant proteins of dPDPN and developed anti-dPDPN mAbs...
August 2016: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/27367430/influence-of-particle-shedding-from-silicone-tubing-on-antibody-stability
#18
Verena Saller, Constanze Hediger, Julia Matilainen, Ulla Grauschopf, Karoline Bechtold-Peters, Hanns-Christian Mahler, Wolfgang Friess
OBJECTIVES: Peristaltic pumps are increasingly employed during fill & finish operations of a biopharmaceutical drug, due to sensitivity of many biological products to rotary piston pump-related stresses. Yet, possibly also unit operations using peristaltic pumps may shed particulates into the final product due to abrasion from the employed tubing. It was the aim of this study to elucidate the potential influence of particles shed from peristaltic pump tubing on the stability of a drug product...
July 1, 2016: Journal of Pharmacy and Pharmacology
https://www.readbyqxmd.com/read/27328060/establishment-of-mouse-monoclonal-antibody-lpmab-13-against-human-podoplanin
#19
Satoshi Ogasawara, Mika K Kaneko, Ryusuke Honma, Hiroharu Oki, Yuki Fujii, Michiaki Takagi, Hiroyoshi Suzuki, Yukinari Kato
Podoplanin (PDPN)/Aggrus is a type-I transmembrane sialoglycoprotein, which possesses a platelet aggregation-stimulating (PLAG) domain. The O-glycosylation on Thr52 of human PDPN (hPDPN) is critical for the interaction of hPDPN with C-type lectin-like receptor-2 (CLEC-2), resulting in platelet aggregation. Many anti-hPDPN monoclonal antibodies (MAbs) against PLAG domains and non-PLAG domains have been established; however, mouse anti-PLAG2/3 MAb, the epitope of which is consistent with rat anti-PLAG2/3 MAb NZ-1, has not been established...
June 2016: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/27325598/assessing-the-risk-of-leachables-from-single-use-bioprocess-containers-through-protein-quality-characterization
#20
Nina Xiao, Colin D Medley, Ian Shieh, Gregory Downing, Shelly Pizarro, Jun Liu, Ankit R Patel
Leachables from single-use bioprocess containers (BPCs) are a source of process-related impurities that have the potential to alter product quality of biotherapeutics and impact patient health. Leachables often exist at very low concentrations making it difficult to detect their presence and challenging to assess their impact on protein quality. A small-scale stress model based on assessing protein stability was developed to evaluate the potential risks associated with storing biotherapeutics in disposable bags caused by the presence of leachables...
June 20, 2016: PDA Journal of Pharmaceutical Science and Technology
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