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https://www.readbyqxmd.com/read/28728084/purification-of-monoclonal-antibodies-entirely-in-flow-through-mode
#1
Tsuyoshi Yamada, Koichi Yamamoto, Takashi Ishihara, Shigeru Ohta
A depth filter is widely used in biopharmaceutical manufacturing process. In this study, we found that buffer exchange to reduce conductivity dramatically improved the removal of impurities in depth filtration. The host cell protein clearance was comparable to that of protein A affinity chromatography, which is generally used as a first capture step in monoclonal antibody purification. In addition, a combination of different depth filters showed enhanced purification. Additional flow-through purification is possible without any sample preparation, and a biopharmaceutical-quality purity level (<100ppm for host cell protein and <5% for high molecular weight species) can be attained...
July 3, 2017: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
https://www.readbyqxmd.com/read/28726668/formulation-and-technology-development-of-herbal-phenolic-biopolymer-containing-films-for-burn-treatment
#2
S Gokadze, V Barbakadze, K Mulkijanyan, A Bakuridze, L Bakuridze
Application of phytofilms based on biosolublepolymers is considered as a prospectivemethod for burn treatment . Herbal remedies contain biologically active substances, that are relatively less toxic, do not cause skin irritation or allergic reactions and, importantly, affectstrains of the microorganisms and viruses resistant to antibiotics and synthetic drugs. Nowadays, the advantages are given to such burn healing drugs, which along with high specific efficacy, have analgesic, anti-inflammatory and antimicrobial effects, and don't irritate the tissues...
June 2017: Georgian Medical News
https://www.readbyqxmd.com/read/28726207/approaches-to-controlled-co-amplification-of-genes-for-production-of-biopharmaceuticals-study-of-the-insertion-and-amplification-dynamics-of-genetic-cassettes-in-the-genome-of-chinese-hamster-ovary-cells-during-co-expression-of-compatible-pair-of-plasmids
#3
S V Kovnir, N A Orlova, Yu А Khodak, M P Kondrashova, A G Gabibov, K G Skryabin, A I Vorobiev, I I Vorobiev
Plasmid vector family p1.1 based on non-coding regions of Chinese hamster housekeeping gene EEF1A and concatemer of Epstein-Barr virus terminal repeat increases the frequency of genome integration and provides rapid amplification of the target genes in the genome. For a pair of fluorescent proteins eGFP and mCherry it was shown that p1.1 vectors bearing dihydrofolate reductase and glutamine synthetase selection markers upon co-transfection into CHO DG44 cell line allow obtaining a polyclonal cell population in which ~70% of cells express both genes...
July 18, 2017: Bulletin of Experimental Biology and Medicine
https://www.readbyqxmd.com/read/28725194/sophisticated-cloning-fermentation-and-purification-technologies-for-an-enhanced-therapeutic-protein-production-a-review
#4
REVIEW
Sanjeev K Gupta, Pratyoosh Shukla
The protein productions strategies are crucial towards the development of application based research and elucidating the novel purification strategies for industrial production. Currently, there are few innovative avenues are studies for cloning, upstream, and purification through efficient bioprocess development. Such strategies are beneficial for industries as well as proven to be vital for effectual therapeutic protein development. Though, these techniques are well documented, but, there is scope of addition to current knowledge with novel and new approaches and it will pave new avenues in production of recombinant microbial and non-microbial proteins including secondary metabolites...
2017: Frontiers in Pharmacology
https://www.readbyqxmd.com/read/28724165/cytogenotoxicity-of-inclusion-complexes-of-diazepam-with-2-hydroxypropyl-%C3%AE-cyclodextrin
#5
Jasmina Hadžiabdić, Nevenka Kopjar, Davor Želježić, Selma Špirtović-Halilović, Davorka Završnik
Background Diazepam, as one of the most frequent prescribed drug from 1,4-benzodiazepine group, has certain limitations in pharmaceutical technology due to its poor solubility in water. By forming inclusion complexes with 2-hydroxypropyl-β-cyclodextrin, diazepam's biopharmaceutical properties can be greatly improved. Aim Aim of this research was to in vitro evaluate genotoxicity of prepared novel complexes of diazepam and their influence on proliferation of human peripheral blood lymphocytes. Methods For identification of possible genotoxicity of diazepam inclusion complexes, cytokinesis-block micronucleus assay has been chosen...
July 19, 2017: Drug Research
https://www.readbyqxmd.com/read/28722325/polymer-mediated-flocculation-of-transient-cho-cultures-as-a-simple-high-throughput-method-to-facilitate-antibody-discovery
#6
Matthew G Schmitt, Yashas Rajendra, Maria D Hougland, Jeffrey S Boyles, Gavin C Barnard
Most biopharmaceutical drugs, especially monoclonal antibodies (mAbs), bispecific antibodies (BsAbs) and Fc-fusion proteins, are expressed using Chinese Hamster Ovary (CHO) cell lines. CHO cells typically yield high product titers and high product quality. Unfortunately, CHO cell lines also generate high molecular weight (HMW) aggregates of the desired product during cell culture along with CHO host cell protein (HCP) and CHO DNA. These immunogenic species, co-purified during Protein A purification, must be removed in a multi-step purification process...
July 19, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28722112/in-silico-and-in-vitro-prediction-of-gastrointestinal-absorption-from-potential-drug-eremantholide-c
#7
Tamires G Caldeira, Dênia A Saúde-Guimarães, André B Dezani, Cristina Helena Dos Reis Serra, Jacqueline de Souza
OBJECTIVES: Analysis of the biopharmaceutical properties of eremantholide C, sesquiterpene lactone with proven pharmacological activity and low toxicity, is required to evaluate its potential to become a drug. METHODS: Preliminary analysis of the physicochemical characteristics of eremantholide C was performed in silico. Equilibrium solubility was evaluated using the shake-flask method, at 37.0 °C, 100 rpm during 72 h in biorelevant media. The permeability was analysed using parallel artificial membrane permeability assay, at 37...
July 19, 2017: Journal of Pharmacy and Pharmacology
https://www.readbyqxmd.com/read/28721831/transient-production-of-recombinant-pharmaceutical-proteins-in-plants-evolution-and-perspectives
#8
Lilya Kopertekh, Joachim Schiemann
During the last two decades the production of pharmaceutical proteins in plants evolved from proof of concept to established technology adopted by several biotechnological companies. This progress is particularly based on intensive research starting stable genetic transformation and moving to transient expression. Due to its advantages in yield and speed of protein production transient expression platforms became leading plant-based manufacturing technology. Current transient expression methods rely on Agrobacterium-mediated delivery of expression vectors into plant cells...
July 18, 2017: Current Medicinal Chemistry
https://www.readbyqxmd.com/read/28721592/construction-purification-and-characterization-of-a-homodimeric-granulocyte-colony-stimulating-factor
#9
Gitana Mickiene, Indre Dalgediene, Zilvinas Dapkunas, Gintautas Zvirblis, Henrikas Pesliakas, Algirdas Kaupinis, Mindaugas Valius, Edita Mistiniene, Milda Pleckaityte
Granulocyte colony-stimulating factor (G-CSF) has found widespread clinical application, and modified forms with improved biopharmaceutical properties have been marketed as well. PEGylation, the covalent modification of G-CSF with polyethylene glycol (PEG), has a beneficial effect on drug properties, but there are concerns connected to the immunogenicity of PEGylated compounds and bioaccumulation of the synthetic polymer. To overcome challenges connected with chemical modifications, we developed fusion proteins composed of two G-CSF molecules connected via different peptide linkers...
July 18, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28720219/monitoring-leachables-from-single-use-bioreactor-bags-for-mammalian-cell-culture-by-dispersive-liquid-liquid-microextraction-followed-by-ultra-high-performance-liquid-chromatography-quadrupole-time-of-flight-mass-spectrometry
#10
N Dorival-García, J Bones
A method for the identification of leachables in chemically defined media for CHO cell culture using dispersive liquid-liquid microextraction (DLLME) and UHPLC-MS is described. A Box-Behnken design of experiments (DoE) approach was applied to obtain the optimum extraction conditions of the target analytes. Performance of DLLME as extraction technique was studied by comparison of two commercial chemically defined media for CHO cell culture. General extraction conditions for any group of leachables, regardless of their specific chemical functionalities can be applied and similar optimum conditions were obtained with the two media...
July 3, 2017: Journal of Chromatography. A
https://www.readbyqxmd.com/read/28718062/update-on-biosimilars-in-asia
#11
REVIEW
Wen-Chan Tsai
PURPOSE OF REVIEW: This review provides an update on the evolution of, manufacturing of, and regulations for biosimilars in Asian countries. RECENT FINDINGS: The use of biologics revolutionized the treatment of various diseases. However, the high cost of biologics remained unaffordable for most Asian patients and increases the financial burden of Asian governments. The development of biosimilars provides the best solution for this predicament. A great boom of biosimilars is developing in Asia...
August 2017: Current Rheumatology Reports
https://www.readbyqxmd.com/read/28716814/potency-matched-dual-cytokine-antibody-fusion-proteins-for-cancer-therapy
#12
Roberto De Luca, Alex Soltermann, Francesca Pretto, Catherine Pemberton-Ross, Giovanni Pellegrini, Sarah Wulhfard, Dario Neri
A novel biopharmaceutical, consisting of the F8 monoclonal antibody (specific to a splice isoform of fibronectin) simultaneously fused to both tumor necrosis factor and interleukin-2, was found to react with the majority of solid tumors and hematological malignancies in mouse and man, but not with healthy adult tissues. The product selectively localized to neoplastic lesions in vivo, as evidenced by quantitative biodistribution studies using radioiodinated protein preparations. When the potency of the cytokine payloads was matched by a single-point mutation, the resulting fusion protein (IL2-F8-TNF(mut)) eradicated soft-tissue sarcomas in immunocompetent mice, which did not respond to individual antibody-cytokine fusion proteins or by standard doxorubicin treatment...
July 17, 2017: Molecular Cancer Therapeutics
https://www.readbyqxmd.com/read/28716374/high-throughput-quantitation-of-fc-containing-recombinant-proteins-in-cell-culture-supernatant-by-fluorescence-polarization-spectroscopy
#13
Ben Thompson, Jerry Clifford, Mike Jenns, Andrew Smith, Ray Field, Kalpana Nayyar, David C James
Measurement of recombinant protein product titer critically underpins all biopharmaceutical manufacturing process development, as well as diverse research and discovery activity. Here, we describe a simple rapid (<2 min per 96 samples) 96-well microplate-based assay that enables high-throughput quantitation of recombinant immunoglobulin G and Fc-containing IgG derivatives in mammalian cell culture supernatant over a wide dynamic range of 2.5-80 mg/L, using microplate fluorescence polarization (FP) spectroscopy...
July 14, 2017: Analytical Biochemistry
https://www.readbyqxmd.com/read/28714211/modeling-of-bispecific-antibody-elution-in-mixed-mode-cation-exchange-chromatography
#14
Yi Feng Lee, Simon Kluters, Mirjam Hillmann, Thomas von Hirschheydt, Christian Frech
The increasing demand for cost-efficient manufacturing of biopharmaceuticals has been the main driving force for the development of novel chromatography resins, which resulted in the development of multimodal or mixed-mode chromatographic resins. Most of them combine electrostatic and hydrophobic functionalities and are designed to deliver unique selectivity and increased binding capacities also at increased ionic strength. However, the mechanism of the protein-resin interaction in mixed-mode chromatography is still not fully understood...
July 17, 2017: Journal of Separation Science
https://www.readbyqxmd.com/read/28711730/purification-and-characterization-of-a-long-acting-ciliary-neurotrophic-factor-via-genetically-fused-with-an-albumin-binding-domain
#15
Longfu Xu, Chun Zhang, Liping Liu, Yao Zhang, Qi Wang, Jian Wang, Yongdong Liu, Zhiguo Su
Ciliary neurotrophic factor (CNTF) is a promising candidate for the treatment of neurodegenerative or metabolic diseases, but suffers rapid clearance in body. Herein we constructed a new long-acting recombinant human CNTF (rhCNTF) by genetic fusion with an albumin-binding domain (ABD) through a flexible peptide linker, hoping to endow the new molecule prolonged serum circulation time by binding with endogenous human serum albumin (HSA) and then utilizing the naturally long-half-life property of HSA. This fused protein rhCNTF-ABD was expressed in Escherichia coli mainly in the soluble form and purified through a two-step chromatography, with purity of 95% and a high yield of 90-100 mg/L culture...
July 12, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28711592/producing-amorphous-solid-dispersions-via-co-precipitation-and-spray-drying-impact-to-physicochemical-and-biopharmaceutical-properties
#16
Amanda K P Mann, Luke Schenck, Athanas Koynov, Alfred C F Rumondor, Xiaoling Jin, Melanie Marota, Chad Dalton
Many small molecule active pharmaceutical ingredients (APIs) exhibit low aqueous solubility, and benefit from generation of amorphous dispersions of the API and polymer to improve their dissolution properties. Spray drying and hot melt extrusion are two common methods to produce these dispersions; however, for some systems these approaches may not be optimal, and it would be beneficial to have an alternative route. Herein, amorphous solid dispersions of Compound A, a low solubility weak acid, and copovidone were made by conventional spray drying and co-precipitation...
July 12, 2017: Journal of Pharmaceutical Sciences
https://www.readbyqxmd.com/read/28710856/quantitative-intracellular-flux-modeling-and-applications-in-biotherapeutic-development-and-production-using-cho-cell-cultures
#17
REVIEW
Zhuangrong Huang, Dong-Yup Lee, Seongkyu Yoon
Chinese hamster ovary (CHO) cells have been widely used for producing many recombinant therapeutic proteins. Constraint-based modeling, such as flux balance analysis (FBA) and metabolic flux analysis (MFA), has been developing rapidly for the quantification of intracellular metabolic flux distribution at a systematic level. Such methods would produce detailed maps of flows through metabolic networks, which contribute significantly to better understanding of metabolism in cells. Although these approaches have been extensively established in microbial systems, their application to mammalian cells is sparse...
July 15, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28710854/opportunities-and-challenges-of-real-time-release-testing-in-biopharmaceutical-manufacturing
#18
REVIEW
Mo Jiang, Kristen Severson, J Christopher Love, Helena Madden, Patrick Swann, Li Zang, Richard D Braatz
Real-time release testing (RTRT) is defined as "the ability to evaluate and ensure the quality of in-process and/or final drug product based on process data, which typically includes a valid combination of measured material attributes and process controls" (ICH Q8(R2)). This article discusses sensors (process analytical technology, PAT) and control strategies that enable RTRT for the spectrum of critical quality attributes (CQAs) in biopharmaceutical manufacturing. Case studies from the small-molecule and biologic pharmaceutical industry are described to demonstrate how RTRT can be facilitated by integrated manufacturing and multivariable control strategies to ensure the quality of products...
July 15, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28710851/generation-of-apoptosis-resistant-hek293-cells-with-crispr-cas-mediated-quadruple-gene-knockout-for-improved-protein-and-virus-production
#19
Weifeng Zhang, Dan Xiao, Linlin Shan, Junli Zhao, Qinwen Mao, Haibin Xia
Apoptosis has important functions during pathophysiologic processes. However, from a biopharmaceutical point of view, active apoptosis of host cells is undesirable during viral packaging or protein expression, because it decreases the efficiency of viral or protein production. Here we used the CRISPR/Cas technique to knock out four pro-apoptotic genes, Caspase3, Caspase6, Caspase7 and AIF1, in HEK293 cells, and successfully produced an apoptosis-resistant cell line. Furthermore, this cell line showed higher expression levels of pro-apoptotic proteins and higher packaging efficiency for the virus carrying these proteins than control HEK293 cells...
July 15, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28710515/quantitative-lc-esi-srm-ms-of-antibody-biopharmaceuticals-use-of-a-homologous-antibody-as-an-internal-standard-and-three-step-method-development
#20
Fumio Osaki, Kenji Tabata, Tomoyuki Oe
Monoclonal antibody-based therapeutic agents (antibody drugs) have attracted considerable attention as a new type of drug. Concomitantly, the use of quantitative approaches for characterizing antibody drugs, such as liquid chromatography (LC)-mass spectrometry (MS), has increased. Generally, selective quantification of antibody drugs is done using unique peptides from variable regions (V H and V L) as surrogate peptides. Further, numerous internal standards (ISs) such as stable isotope-labeled (SIL)-intact proteins and SIL-surrogate peptides are used...
July 14, 2017: Analytical and Bioanalytical Chemistry
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