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https://www.readbyqxmd.com/read/29110285/chemical-and-biophysical-characteristics-of-monoclonal-antibody-solutions-containing-aggregates-formed-during-metal-catalyzed-oxidation
#1
Linda O Narhi, Quanzhou Luo, Jette Wypych, Riccardo Torosantucci, Andrea Hawe, Kiyoshi Fujimori, Yasser Nashed-Samuel, Vibha Jawa, Marisa K Joubert, Wim Jiskoot
PURPOSE: To physicochemically characterize and compare monoclonal antibody (mAb) solutions containing aggregates generated via metal catalyzed oxidation (MCO). METHODS: Two monoclonal IgG2s (mAb1 and mAb2) and one monoclonal IgG1 (rituximab) were exposed to MCO with the copper/ascorbic acid oxidative system, by using several different methods. The products obtained were characterized by complementary techniques for aggregate and particle analysis (from oligomers to micron sized species), and mass spectrometry methods to determine the residual copper content and chemical modifications of the proteins...
November 6, 2017: Pharmaceutical Research
https://www.readbyqxmd.com/read/29090969/expression-of-cat-podoplanin-in-feline-squamous-cell-carcinomas
#2
Shunsuke Itai, Shinji Yamada, Mika K Kaneko, Hiroyuki Harada, Yumiko Kagawa, Satoru Konnai, Yukinari Kato
Oral squamous cell carcinoma is an aggressive tumor in cats; however, molecular-targeted therapies against this tumor, including antibody therapy, have not been developed. Sensitive and specific monoclonal antibodies (mAbs) against highly expressed membrane proteins are needed to develop antibody therapies. Podoplanin, a type I transmembrane glycoprotein, is expressed in many human malignant tumors, including brain tumor, esophageal cancer, lung cancer, mesothelioma, and oral cancer. Podoplanin binds to C-type lectin-like receptor-2 (CLEC-2) and activates platelet aggregation, which is involved in cancer metastasis...
November 1, 2017: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/29055527/multi-state-steric-mass-action-model-and-case-study-on-complex-high-loading-behavior-of-mab-on-ion-exchange-tentacle-resin
#3
Juliane Diedrich, William Heymann, Samuel Leweke, Stephen Hunt, Robert Todd, Christian Kunert, Will Johnson, Eric von Lieres
Tentacle resins for IEX are increasingly applied in preparative chromatography for their higher selectivity and higher capacities in comparison to IEX resins without tentacles. However, tentacle resins are often observed to cause unusual elution behavior of monoclonal antibodies under high loading conditions. Understanding this elution behavior is important for a quality by design approach, as it is now mandated by regulatory agencies. A model-based analysis of load, wash and gradient elution is performed for a monoclonal antibody (mAb) on Fractogel SO3(-)...
September 20, 2017: Journal of Chromatography. A
https://www.readbyqxmd.com/read/29054386/interlaced-size-exclusion-chromatography-for-faster-protein-analysis
#4
Julia Kahle, Hermann Wätzig
In this research note, interlaced Size Exclusion Chromatography (SEC) is used to shorten the duration of protein analysis. The applied method has a threefold higher sample throughput than comparable ones using conventional SEC. Further important advantages are the identical area's precision and the preservation of protein stability. Two different applications are presented where a short time interval of analysis is very important. One application is the determination of aggregates of monoclonal antibodies (mAbs)...
October 17, 2017: European Journal of Pharmaceutics and Biopharmaceutics
https://www.readbyqxmd.com/read/29037466/the-use-of-a-groel-bli-biosensor-to-rapidly-assess-pre-aggregate-populations-for-antibody-solutions-exhibiting-different-stability-profiles
#5
Samantha E Pace, Sangeeta B Joshi, Reza Esfandiary, Robert Stadelman, Steven M Bishop, C R Middaugh, Mark Fisher, David B Volkin
An automated method using biotinylated GroEL-streptavidin biosensors with Bio-Layer Interferometry (GroEL-BLI) was evaluated to detect the formation of transiently formed, pre-aggregate species in various pharmaceutically relevant monoclonal antibody (mAb) samples. The relative aggregation propensity of various IgG1 and IgG4 mAbs was rank-ordered using the GroEL-BLI biosensor method, and the least stable IgG4 mAb was subjected to different stresses including elevated temperatures, acidic pH, and addition of guanidine-HCl...
October 13, 2017: Journal of Pharmaceutical Sciences
https://www.readbyqxmd.com/read/28987959/reanalysis-of-the-role-of-pronase-treatment-of-b-cells-in-the-flow-cytometric-crossmatch-assay-fc-receptor-is-not-the-primary-target
#6
Nicholas K Brown, James R Meade, Jinguo Wang, Susana R Marino
Pronase, a mixture of nonspecific bacterial proteases, is used to pretreat human lymphocytes to prevent false-positive B cell results in the flow cytometric crossmatch (FCXM) assay. The target of pronase has been reported to be B cell-expressed Fc receptors, which nonspecifically bind IgG. As pronase use in FCXM can induce other complications, including degradation of HLA leading to inappropriate FCXM results, and false-positive T cell results when testing serum from HIV-positive patients, we tested whether specifically blocking Fc receptor CD32 could replace pronase...
October 5, 2017: Human Immunology
https://www.readbyqxmd.com/read/28986999/one-step-affinity-capture-and-precipitation-for-improved-purification-of-an-industrial-monoclonal-antibody-using-z-elp-functionalized-nanocages
#7
Andrew R Swartz, Xuankuo Xu, Steven J Traylor, Zheng J Li, Wilfred Chen
Protein A chromatography has been identified as a potential bottleneck in the monoclonal antibody production platform, leading to increased interest in non-chromatographic capture technologies. Affinity precipitation using environmentally responsive, Z-domain-elastin-like polypeptide (Z-ELP) fusion proteins has been shown to be a promising alternative. However, elevated temperature and salt concentrations necessary for precipitation resulted in decreased antibody monomer content and reduced purification capacity...
October 7, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28946123/characterization-of-30-therapeutic-antibodies-and-related-products-by-size-exclusion-chromatography-feasibility-assessment-for-future-mass-spectrometry-hyphenation
#8
Alexandre Goyon, Valentina D'Atri, Olivier Colas, Szabolcs Fekete, Alain Beck, Davy Guillarme
Despite the popularity of targeted and immune therapies, the number of studies dealing with the quantitation of aggregates for Food and Drug Administration (FDA) and European Medicines Agency (EMA) approved mAb and related products are still very scarce in literature. In this work, 30 therapeutic proteins including monoclonal antibodies (mAbs), antibody-drug conjugates (ADCs), Fc-fusion proteins and a bi-specific antibody (bsAb) were investigated using size exclusion chromatography (SEC). Their levels of high molecular weight species (HMWS) were experimentally estimated between 0...
October 15, 2017: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
https://www.readbyqxmd.com/read/28945165/dengue-virus-and-its-relation-to-human-glycoprotein-iib-iiia-revealed-by-fluorescence-microscopy-and-flow-cytometry
#9
Nattapol Attatippaholkun, Yaowalak U-Pratya, Panthipa Supraditaporn, Chanchao Lorthongpanich, Kovit Pattanapanyasat, Surapol Issaragrisil
Understanding dengue virus (DENV)-induced hemorrhage remains a challenging jigsaw puzzle with many pieces missing to understand the complex interactions between DENV and blood coagulation system. To use flow cytometry studying the interactions between DENV and human platelet aggregation receptor, glycoprotein IIb/IIIa (gpIIb/IIIa), directly conjugated fluorochrome monoclonal antibody (mAb) is essential to facilitate multifluorochrome immunostaining. However, the obstacle was that no directly conjugated fluorochrome-anti-DENV mAb had been commercially available...
November 2017: Viral Immunology
https://www.readbyqxmd.com/read/28938559/doxorubicin-loaded-platelets-conjugated-with-anti-cd22-mabs-a-novel-targeted-delivery-system-for-lymphoma-treatment-with-cardiopulmonary-avoidance
#10
Peipei Xu, Huaqin Zuo, Rongfu Zhou, Fan Wang, Xu Liu, Jian Ouyang, Bing Chen
B-cell lymphoma accounts for approximately 85% of all adult non-Hodgkin's lymphoma cases. Doxorubicin (DOX) is an indispensable drug for the treatment of non-Hodgkin's lymphoma. However, DOX causes severe cardiotoxicity, which limits its use in conventional treatment strategies. In this study, we developed a novel drug delivery system for lymphoma treatment: DOX-loaded platelets that were conjugated with anti-CD22 monoclonal antibodies (mAbs) (DOX-platelet-CD22). Platelets are bio- and immune-compatible drug carriers that can prolong the circulation time of drugs...
August 29, 2017: Oncotarget
https://www.readbyqxmd.com/read/28935894/strain-dependent-neutralization-reveals-antigenic-variation-of-human-parechovirus-3
#11
Eveliina Karelehto, Sabine van der Sanden, James A Geraets, Aušra Domanska, Lonneke van der Linden, Dionne Hoogendoorn, Gerrit Koen, Hetty van Eijk, Shabih Shakeel, Tim Beaumont, Menno de Jong, Dasja Pajkrt, Sarah J Butcher, Katja C Wolthers
Human parechovirus 3 (HPeV3), a member of the Picornavirus family, is frequently detected worldwide. However, the observed seropositivity rates for HPeV3 neutralizing antibodies (nAbs) vary from high in Japan to low in the Netherlands and Finland. To study if this can be explained by technical differences or antigenic diversity among HPeV3 strains included in the serological studies, we determined the neutralizing activity of Japanese and Dutch intravenous immunoglobulin batches (IVIG), a rabbit HPeV3 hyperimmune polyclonal serum, and a human HPeV3-specific monoclonal antibody (mAb) AT12-015, against the HPeV3 A308/99 prototype strain and clinical isolates from Japan, the Netherlands and Australia, collected between 1989 and 2015...
September 21, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28881096/integrated-chromatographic-platform-for-simultaneous-separation-of-charge-variants-and-aggregates-from-monoclonal-antibody-therapeutic-products
#12
Nikhil Kateja, Devashish Kumar, Avinash Godara, Vijesh Kumar, Anurag S Rathore
Achieving consistent product quality of a biotherapeutic is a major target for any biopharmaceutical manufacturer, even more for a biosimilar producer as comparability with the innovator product is a regulatory expectation. The complexity of biotherapeutic products and their tedious manufacturing processes, however, make this a non-trivial exercise. The primary motivation of this work is to develop an integrated chromatographic platform for purification of monoclonal antibody (mAb) therapeutics that can deliver the desired separation of both charge variants and aggregates, in addition to the process related impurities like host cell proteins (HCP) and host cell DNA...
September 7, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28880521/peptide-dendrons-as-thermal-stability-amplifiers-for-immunoglobulin-g1-monoclonal-antibody-biotherapeutics
#13
Rohit Bansal, Sameer Dhawan, Soumili Chattopadhyay, Govind P Maurya, V Haridas, Anurag S Rathore
Biotherapeutics such as monoclonal antibodies (mAbs) have a major share of the pharmaceutical industry for treatment of life-threatening chronic diseases such as cancer, skin ailments, and immune disorders. Instabilities such as aggregation, fragmentation, oxidation, and reduction have resulted in the practice of storing these products at low temperatures (-80 to -20 °C). However, reliable storage at these temperatures can be a challenge, particularly in developing and underdeveloped countries; hence, lately, there has been a renewed interest in creating formulations that would offer stability at higher temperatures (25 to 55 °C)...
October 18, 2017: Bioconjugate Chemistry
https://www.readbyqxmd.com/read/28862080/evaluation-of-brain-targeted-chitosan-nanoparticles-through-blood-brain-barrier-cerebral-microvessel-endothelial-cells
#14
Adem Sahin, Digdem Yoyen-Ermis, Secil Caban-Toktas, Utku Horzum, Yesim Aktas, Patrick Couvreur, Gunes Esendagli, Yilmaz Capan
The blood-brain barrier (BBB) is the major problem for the treatment of central nervous system diseases. A previous study from our group showed that the brain-targeted chitosan nanoparticles-loaded with large peptide moieties can rapidly cross the barrier and provide neuroprotection. The present study aims to determine the efficacy of the brain-targeted chitosan nanoparticles' uptake by the human BBB cerebral microvessel endothelial cells (hCMECs) and to investigate the underlying mechanisms for enhanced cellular entry...
September 13, 2017: Journal of Microencapsulation
https://www.readbyqxmd.com/read/28843351/effect-of-polysorbate-20-and-polysorbate-80-on-the-higher-order-structure-of-a-monoclonal-antibody-and-its-fab-and-fc-fragments-probed-using-2d-nuclear-magnetic-resonance-spectroscopy
#15
Surinder M Singh, Swati Bandi, David N M Jones, Krishna M G Mallela
We examined how polysorbate 20 (PS20; Tween 20) and polysorbate 80 (PS80; Tween 80) affect the higher-order structure of a monoclonal antibody (mAb) and its antigen-binding (Fab) and crystallizable (Fc) fragments, using near-UV circular dichroism and 2D nuclear magnetic resonance (NMR). Both polysorbates bind to the mAb with submillimolar affinity. Binding causes significant changes in the tertiary structure of mAb with no changes in its secondary structure. 2D (13)C-(1)H methyl NMR indicates that with increasing concentration of polysorbates, the Fab region showed a decrease in crosspeak volumes...
August 24, 2017: Journal of Pharmaceutical Sciences
https://www.readbyqxmd.com/read/28815933/an-%C3%AE-iib-%C3%AE-3-antagonist-prevents-thrombosis-without-causing-fc-receptor-%C3%AE-chain-iia-mediated-thrombocytopenia
#16
Y-J Kuo, Y-R Chen, C-C Hsu, H-C Peng, T-F Huang
Essentials FcγRIIa-mediated thrombocytopenia is associated with drug-dependent antibodies (DDAbs). We investigated the correlation between αIIb β3 binding epitopes and induction of DDAbs. An FcγRIIa-transgenic mouse model was used to evaluate thrombocytopenia among anti-thrombotics. An antithrombotic with binding motif toward αIIb β-propeller domain has less bleeding tendency. SUMMARY: Background Thrombocytopenia, a common side effect of Arg-Gly-Asp-mimetic antiplatelet drugs, is associated with drug-dependent antibodies (DDAbs) that recognize conformation-altered integrin αIIb β3 ...
August 16, 2017: Journal of Thrombosis and Haemostasis: JTH
https://www.readbyqxmd.com/read/28776673/molecular-characterization-of-excipients-preferential-interactions-with-therapeutic-monoclonal-antibodies
#17
Jehoon Kim, Mark R H Krebs, Bernhardt L Trout
OBJECTIVES: This study reports global and local monoclonal antibody (mAb)-excipient interactions and the resulting thermodynamic and stabilization effects. METHODS: Molecular dynamics simulations are applied to quantify the interactions of key excipients (sucrose, sorbitol, arginine, citrate, histidine and NaCl) used in the formulations of three mAbs. The dynamic surface properties of the mAbs and preferential interaction coefficients for the excipients based on validated potential parameters are computed, in addition to the spatial aggregation propensity...
August 4, 2017: Journal of Pharmacy and Pharmacology
https://www.readbyqxmd.com/read/28758834/preferential-interactions-of-trehalose-l-arginine-hcl-and-sodium-chloride-with-therapeutically-relevant-igg1-monoclonal-antibodies
#18
Chaitanya Sudrik, Theresa Cloutier, Phuong Pham, Hardeep S Samra, Bernhardt L Trout
Preferential interactions of weakly interacting formulation excipients govern their effect on the equilibrium and kinetics of several reactions of protein molecules in solution. Using vapor pressure osmometry, we characterized the preferential interactions of commonly used excipients trehalose, L-arginine.HCl and NaCl with three therapeutically-relevant, IgG1 monoclonal antibodies that have similar size and shape, but differ in their surface hydrophobicity and net charge. We further characterized the effect of these excipients on the reversible self-association, aggregation and viscosity behavior of these antibody molecules...
October 2017: MAbs
https://www.readbyqxmd.com/read/28754261/elucidating-the-weak-protein-protein-interaction-mechanisms-behind-the-liquid-liquid-phase-separation-of-a-mab-solution-by-different-types-of-additives
#19
Guoliang Wu, Shujing Wang, Zhou Tian, Ning Zhang, Han Sheng, Weiguo Dai, Feng Qian
Liquid-liquid phase separation (LLPS) has long been observed during the physical stability investigation of therapeutic protein formulations. The buffer conditions and the presence of various excipients are thought to play important roles in the formulation development of monoclonal antibodies (mAbs). In this study, the effects of several small-molecule excipients (histidine, alanine, glycine, sodium phosphate, sodium chloride, sorbitol and sucrose) with diverse physical-chemical properties on LLPS of a model IgG1 (JM2) solutions were investigated by multiple techniques, including UV-vis spectroscopy, circular dichroism, differential scanning calorimetry/fluorimetry, size exclusion chromatography and dynamic light scattering...
November 2017: European Journal of Pharmaceutics and Biopharmaceutics
https://www.readbyqxmd.com/read/28737447/pmab-52-specific-and-sensitive-monoclonal-antibody-against-cat-podoplanin-for-immunohistochemistry
#20
Shinji Yamada, Shunsuke Itai, Takuro Nakamura, Miyuki Yanaka, Noriko Saidoh, Yao-Wen Chang, Saori Handa, Hiroyuki Harada, Yumiko Kagawa, Osamu Ichii, Satoru Konnai, Mika K Kaneko, Yukinari Kato
Podoplanin (PDPN) is expressed in several normal tissues, such as lymphatic endothelial cells, podocytes of renal glomerulus, and type I alveolar cells of lung. PDPN activates platelet aggregation by binding to C-type lectin-like receptor-2 (CLEC-2) on platelet. Although monoclonal antibodies (mAbs) against human PDPN, mouse PDPN, rat PDPN, rabbit PDPN, dog PDPN, and bovine PDPN have been established, anticat PDPN (cPDPN) mAbs have not been developed. In this study, we immunized mice with Chinese hamster ovary (CHO)-K1 cell lines expressing cPDPN, and developed anti-cPDPN mAbs...
October 2017: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
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