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https://www.readbyqxmd.com/read/28708446/elisa-reagent-coverage-evaluation-by-affinity-purification-tandem-mass-spectrometry
#1
Scott M Henry, Elissa Sutlief, Oscar Salas-Solano, John Valliere-Douglass
Host cell proteins (HCPs) must be adequately removed from recombinant therapeutics by downstream processing to ensure patient safety, product quality, and regulatory compliance. HCP process clearance is typically monitored by enzyme-linked immunosorbent assay (ELISA) using a polyclonal reagent. Recently, mass spectrometry (MS) has been utilized to identify specific HCP process impurities and monitor their clearance. Despite this capability, ELISA remains the preferred analytical approach due to its simplicity and throughput...
July 14, 2017: MAbs
https://www.readbyqxmd.com/read/28692328/fabs-in-tandem-immunoglobulin-is-a-novel-and-versatile-bispecific-design-for-engaging-multiple-therapeutic-targets
#2
Shiyong Gong, Fang Ren, Danqing Wu, Xuan Wu, Chengbin Wu
In recent years, the development of bispecific antibody (bsAb) has become a major trend in the biopharmaceutical industry. By simultaneously engaging two molecular targets, bsAbs show unique mechanisms of action that could lead to clinical benefits unattainable by conventional monoclonal antibodies. Various bsAb generation formats have been described, and several are being investigated in clinical development. However, some bsAb constructs have proven to be problematic due to their unfavorable physicochemical and pharmacokinetic properties, as well as poor manufacturing efficiencies...
July 10, 2017: MAbs
https://www.readbyqxmd.com/read/28682148/pharmacokinetics-of-an-intracerebroventricularly-administered-antibody-in-rats
#3
Yuki Noguchi, Motohiro Kato, Kazuhisa Ozeki, Masaki Ishigai
The pharmacokinetics (PK) of an antibody in the brain and the spinal cord is insufficiently understood, which is an obstacle to the discovery of antibody drugs that target diseases in the central nervous system. In this study, we focused on the elimination of IgG from cerebrospinal fluid (CSF) circulating in the brain and the spinal cord in rats, and, in order to evaluate the influence of CSF bulk flow on the clearance of IgG, also examined the PK of inulin in CSF. To monitor their concentrations in CSF, IgG and inulin were co-administered into the lateral ventricle via a catheter, and CSF was collected from the cisterna magna via another catheter time-sequentially...
July 6, 2017: MAbs
https://www.readbyqxmd.com/read/28678617/safety-of-biologics-therapy-monoclonal-antibodies-cytokines-fusion-proteins-hormones-enzymes-coagulation-proteins-vaccines-botulinum-toxins
#4
https://www.readbyqxmd.com/read/28657418/chemically-defined-camelid-antibody-bioconjugate-for-the-magnetic-resonance-imaging-of-alzheimer-s-disease
#5
Matthias Vandesquille, Tengfei Li, Chrystelle Po, Christelle Ganneau, Pascal Lenormand, Clémence Dudeffant, Christian Czech, Fiona Grueninger, Charles Duyckaerts, Benoît Delatour, Marc Dhenain, Pierre Lafaye, Sylvie Bay
Today, molecular imaging of neurodegenerative diseases is mainly based on small molecule probes. Alternatively, antibodies are versatile tools that may be developed as new imaging agents. Indeed, they can be readily obtained to specifically target any antigen of interest and their scaffold can be functionalized. One of the critical issues involved in translating antibody-based probes to the clinic is the design and synthesis of perfectly-defined conjugates. Camelid single-domain antibody-fragments (VHHs) are very small and stable antibodies that are able to diffuse in tissues and potentially cross the blood brain barrier (BBB)...
June 28, 2017: MAbs
https://www.readbyqxmd.com/read/28640663/glycosylation-profile-and-biological-activity-of-remicade%C3%A2-compared-with-flixabi%C3%A2-and-remsima%C3%A2
#6
Changsoo Lee, Min Jeong, JongAh Joanne Lee, Saebom Seo, Sung Chun Cho, Wei Zhang, Orlando Jaquez
As biosimilars enter the market, comparisons of product quality are needed. Manufacturing differences may lead to differences in critical quality attributes, which affect efficacy. Therefore, critical quality attributes (structure and biological activity) of Remicade® and of 2 biosimilar products (Flixabi®/Renflexis® and Remsima®/Inflectra®) were determined. We assessed binding to tumor necrosis factor in a fluorescence competitive binding assay; potency in a luciferase reporter gene assay; percentages of galactosylated glycan, afucose plus high mannosylated glycans, and charged glycan; FcγRIIIa (CD16) binding (assessed by 3 methods); and antibody-dependent cell-mediated cytotoxicity (ADCC) in the NK92-CD16a cell line and in peripheral blood mononuclear cells (PBMC)...
June 22, 2017: MAbs
https://www.readbyqxmd.com/read/28621572/changes-to-international-non-proprietary-names-for-antibody-therapeutics-2017-and-beyond-of-mice-men-and-more%C3%A2
#7
Paul W H I Parren, Paul J Carter, Andreas Plückthun
Active pharmaceutical substances require an International Nonproprietary Name (INN)assigned by the World Health Organization (WHO) to obtain market authorization as a medicinal product. INNsare selected to represent a unique, generic namefor a drug enablingunambiguous identification by stakeholders worldwide. INNs may be requested after initiating clinical development of an investigational drug. Pharmaceutical classes are indicated by a common stem or suffix. Currently, INNsfor monoclonal antibody-based drugs are recognized by the suffix, -mab,preceded by a source infixsuch as-xi- (chimeric),-zu- (humanized) or -u- (human) designating the species from which the antibody was derived...
June 16, 2017: MAbs
https://www.readbyqxmd.com/read/28617076/characterization-of-highly-concentrated-antibody-solution-a-toolbox-for-the-description-of-protein-long-term-solution-stability
#8
Marie-Therese Schermeyer, Anna K Wöll, Bas Kokke, Michel Eppink, Jürgen Hubbuch
High protein titers are gaining importance in biopharmaceutical industry. A major challenge in the development of highly concentrated mAb solutions is their long-term stability and often incalculable viscosity. The complexity of the molecule itself, as well as the various molecular interactions, make it difficult to describe their solution behavior. To study the formulation stability, long- and short-range interactions and the formation of complex network structures have to be taken into account. For a better understanding of highly concentrated solutions, we combined established and novel analytical tools to characterize the effect of solution properties on the stability of highly concentrated mAb formulations...
June 15, 2017: MAbs
https://www.readbyqxmd.com/read/28613103/quantification-of-igg-monoclonal-antibody-clearance-in-tissues
#9
Miro J Eigenmann, Ludivine Fronton, Hans Peter Grimm, Michael B Otteneder, Ben-Fillippo Krippendorff
Monoclonal antibodies are an important therapeutic entity, and knowledge of antibody pharmacokinetics has steadily increased over the years. Despite this effort, little is known about the extent of IgG antibody degradation in different tissues of the body. While studies have been published identifying sites of degradation with the use of residualizing and non-residualizing radiolabels, quantitative tissue clearances have not yet been derived. Here, we show that in physiologically-based pharmacokinetic (PBPK) models we can combine mouse data of Indium-111 and Iodine-125 labeled antibodies with prior physiological knowledge to determine tissue-specific intrinsic clearances...
June 14, 2017: MAbs
https://www.readbyqxmd.com/read/28613102/a-high-throughput-platform-for-population-reformatting-and-mammalian-expression-of-phage-display-libraries-to-enable-functional-screening-as-full-length-igg
#10
Xiaodong Xiao, Julie A Douthwaite, Yan Chen, Ben Kemp, Sara Kidd, Jennifer Percival-Alwyn, Alison Smith, Kate Goode, Bonnie Swerdlow, David Lowe, Herren Wu, William F Dall'Acqua, Partha S Chowdhury
Phage display antibody libraries are a rich resource for discovery of potential therapeutic antibodies. Single-chain variable fragment (scFv) libraries are the most common format due to the efficient display of scFv by phage particles and the ease by which soluble scFv antibodies can be expressed for high-throughput screening. Typically, a cascade of screening and triaging activities are performed, beginning with the assessment of large numbers of E. coli-expressed scFv, and progressing through additional assays with individual reformatting of the most promising scFv to full-length IgG...
June 14, 2017: MAbs
https://www.readbyqxmd.com/read/28598281/design-development-and-characterization-of-act017-a-humanized-fab-that-blocks-platelet-s-glycoprotein-vi-function-without-causing-bleeding-risks
#11
Kristell Lebozec, Martine Jandrot-Perrus, Gilles Avenard, Olivier Favre-Bulle, Philippe Billiald
Glycoprotein VI is a platelet-specific collagen receptor critical for in vivo formation of arterial thrombosis. It is also considered as an attractive target for the development of anti-thrombotic drugs because blocking glycoprotein (GP)VI inhibits platelet aggregation without inducing detrimental effects on physiological hemostasis. Here, we present data on the identification, in vitro and ex vivo pharmacology of a humanized Fab fragment designated as ACT017. ACT017 was selected out of 15 humanized variants based upon structural and functional properties...
June 9, 2017: MAbs
https://www.readbyqxmd.com/read/28590212/evidence-for-intermolecular-domain-exchange-in-the-fab-domains-of-dimer-and-oligomers-of-an-igg1-monoclonal-antibody
#12
Yin Luo, Stephen W Raso, Judith Gallant, Colleen Steinmeyer, Yasuko Mabuchi, Zhaojiang Lu, Clifford Entrican, Jason C Rouse
Recombinant protein therapeutics have become increasingly useful in combating human diseases, such as cancer and those of genetic origin. One quality concern for protein therapeutics is the content and the structure of the aggregated proteins in the product, due to the potential immunogenicity of these aggregates. Collective efforts have led to a better understanding of some types of protein aggregates, and have revealed the diversity in the structure and cause of protein aggregation. In this work we used a broad range of analytical techniques to characterize the quinary structure (complexes in which each composing unit maintains native quaternary structure) of the stable non-covalent dimer and oligomers of a monoclonal IgG1λ antibody...
June 7, 2017: MAbs
https://www.readbyqxmd.com/read/28590201/improved-sequence-variant-analysis-strategy-by-automated-false-positive-removal
#13
Wenzhou Li, Jette Wypych, Robert J Duff
Sequence variant analysis (SVA) is critical in therapeutic protein development because it ensures the absence of genetic mutations of a production clone or high-level misincorporations during cell culture. While software for searching sequence variants from mass spectrometry data is available, effectively distinguishing true positives from a large number of false positives in the reported hits or identifications found in the error tolerant search mode is a challenge. This verification process must be done manually and can take several days or even weeks to accomplish...
June 7, 2017: MAbs
https://www.readbyqxmd.com/read/28590151/characterization-of-a-novel-modification-of-a-cho-produced-mab-evidence-for-the-presence-of-tyrosine-sulfation
#14
Jia Zhao, Jason Saunders, Svetlana Dukleska Schussler, Sandra Rios, Francis Kobina Insaidoo, Aleksandr L Fridman, Huijuan Li, Yan-Hui Liu
Herein we describe the investigation of a Chinese hamster ovary (CHO)-expressed human mAb molecule found partially modified by a +80 Da adduct. This mass difference, suggestive of a single sulfation or phosphorylation addition, was observed by mass analysis of the intact and reduced molecule by mass spectrometry (MS). The modification was located on tyrosine 31 (Y31) of the light chain in the complementarity-determining region 1 by liquid chromatography (LC)-MS peptide mapping and electron transfer dissociation fragmentation...
June 7, 2017: MAbs
https://www.readbyqxmd.com/read/28585908/tuning-the-specificity-of-a-two-in-one-fab-against-three-angiogenic-antigens-by-fully-utilizing-the-information-of-deep-mutational-scanning
#15
Patrick Koenig, Sarah Sanowar, Chingwei V Lee, Germaine Fuh
Monoclonal antibodies developed for therapeutic or diagnostic purposes need to demonstrate highly defined binding specificity profiles. Engineering of an antibody to enhance or reduce binding to related antigens is often needed to achieve the desired biological activity without safety concern. Here, we describe a deep sequencing-aided engineering strategy to fine-tune the specificity of an angiopoietin-2 (Ang2)/vascular endothelial growth factor (VEGF) dual action Fab, 5A12.1 for the treatment of age-related macular degeneration...
June 6, 2017: MAbs
https://www.readbyqxmd.com/read/28581887/impact-of-s-sulfocysteine-on-fragments-and-trisulfide-bond-linkages-in-monoclonal-antibodies
#16
Ronja Seibel, Sandra Maier, Alisa Schnellbaecher, Susanne Bohl, Maria Wehsling, Anne Zeck, Aline Zimmer
The quality of recombinant proteins such as monoclonal antibodies produced using Chinese hamster ovary cell-based mammalian systems is dependent on many factors, including cell line, process and cell culture media. Due to these factors, the generated product is heterogeneous and may have chemically-induced modifications or post-translational modifications that affect antibody stability, functionality and, in some cases, patient safety. This study demonstrates that S-sulfocysteine, a cysteine derivative, can increase the antibody specific productivity in different cell lines cultivated with different processes while minimizing trisulfide linkages in generated mAbs, mainly between heavy and light chain...
June 5, 2017: MAbs
https://www.readbyqxmd.com/read/28581886/enhanced-immunization-techniques-to-obtain-highly-specific-monoclonal-antibodies
#17
Rodrigo de Almeida, Cecília Naomi Nakamura, Marina de Lima Fontes, Elenice Deffune, Sérgio Luis Felisbino, Ramon Kaneno, Wagner José Fávaro, Athanase Billis, Marcel Otávio Cerri, Ana Marisa Fusco-Almeida, Maria José Mendes-Giannini, Andrei Moroz
Despite fast advances in genomics and proteomics, monoclonal antibodies (mAbs) are still a valuable tool for areas such as the evolution of basic research in stem cells and cancer, for immunophenotyping cell populations, diagnosing and prognosis of diseases, and for immunotherapy. To summarize different subtractive immunization approaches successfully employed for the production of highly specific antibodies, we identified scientific articles in NCBI PubMed using the following search terms: subtractive immunization, monoclonal antibody, tolerization, neonatal, high-zone tolerance, masking immunization...
June 5, 2017: MAbs
https://www.readbyqxmd.com/read/28581883/discovery-and-characterization-of-a-novel-humanized-anti-il-15-antibody-and-its-relevance-for-the-treatment-of-refractory-celiac-disease-and-eosinophilic-esophagitis
#18
Alain P Vicari, Alain M Schoepfer, Bertrand Meresse, Laurence Goffin, Olivier Léger, Soheila Josserand, Nicolas Guégan, Shida Yousefi, Alex Straumann, Nadine Cerf-Bensussan, Hans-Uwe Simon, Yolande Chvatchko
Interleukin-15 (IL-15) is a critical regulator of immune responses, especially at mucosal interfaces within the gastro-intestinal tract. Here, we describe the discovery and characterization of a humanized antibody to IL-15. Data from its epitope and mode of action, cell biology and primate pharmacology, as well as translational studies in human samples and in vivo proof-of-concept experiments in mouse models demonstrate the therapeutic potential of this new antibody targeting IL-15 for refractory celiac disease and eosinophilic esophagitis...
June 5, 2017: MAbs
https://www.readbyqxmd.com/read/28557665/contribution-of-physiologists-to-the-identification-of-the-humoral-component-of-immunity-in-the-19th-century
#19
Yves-Marie Lahaie, Hervé Watier
The history of antimicrobial humoral immunity usually focuses on the works of the German school at the end of the 19th century, born in the tradition of chemistry and disinfection. Starting from an old quarrel of priority about serotherapy between Emil von Behring (1854-1917) and the French physiologists Charles Richet (1850-1935) and Jules Héricourt (1850-1938), we first confirm that the latter stated the principle of serotherapy in 1888 and put it into practice before the seminal Behring's article in 1890, observing several adverse effects of this new immunotherapy...
May 30, 2017: MAbs
https://www.readbyqxmd.com/read/28541812/engineering-humanized-antibody-framework-sequences-for-optimal-site-specific-conjugation-of-cytotoxins
#20
Jared L Spidel, Earl F Albone, Xin Cheng, Benjamin Vaessen, Sara Jacob, Andrew Z Milinichik, Arielle Verdi, J Bradford Kline, Luigi Grasso
The prevailing techniques used to generate antibody-drug conjugates (ADCs) involve random conjugation of the linker-drug to multiple lysines or cysteines in the antibody. Engineering natural and non-natural amino acids into an antibody has been demonstrated to be an effective strategy to produce homogeneous ADC products with defined drug-to-antibody ratios. We recently reported an efficient residue-specific conjugation technology (RESPECT) where thiol-reactive payloads can be efficiently conjugated to a native unpaired cysteine in position 80 (C80) of rabbit light chains...
May 25, 2017: MAbs
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