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https://www.readbyqxmd.com/read/29714020/epitope-binning-of-human-monoclonal-antibodies-in-classical-sandwich-and-in-tandem-orientation-using-the-octet-system-based-on-biolayer-interferometry
#1
Lisette Bronswijk-Deddens
Epitope binning is a technique used to cluster different monoclonal antibodies by the specific region on the antigen that is recognized by the antibody. This is used to increase the likelihood of choosing lead antibodies with the desired biological activity by selecting antibodies from distinct bins, to confirm that biosimilars or biobetters bind to similar epitopes and to select reagents for diagnostic sandwich and/or ELISA-type assays. This protocol describes epitope binning of seven human monoclonal antibodies to a monovalent antigen in two orthogonal orientations; a classical sandwich and an in-tandem orientation using an eight-channel Octet system...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29551638/an-oxidation-resistant-recombinant-alpha-1-antitrypsin-produced-in-nicotiana-benthamiana
#2
David Z Silberstein, Kalimuthu Karuppanan, Hnin Hnin Aung, Ching-Hsien Chen, Carroll E Cross, Karen A McDonald
Proteases and reactive oxygen species (ROS) have long been implicated in playing key roles in host tissue injury at sites of inflammatory processes dominated by macrophage activations and/or neutrophil infiltrations. Imbalances between proteases/antiproteases and ROS/antioxidants are recognized to contribute to amplification of inflammatory-based host tissue injury. This has been especially well-documented in such respiratory tract diseases as chronic obstructive pulmonary disease, cystic fibrosis, and acute respiratory distress syndrome...
March 15, 2018: Free Radical Biology & Medicine
https://www.readbyqxmd.com/read/29385073/implementation-of-glycan-remodeling-to-plant-made-therapeutic-antibodies
#3
Lindsay D Bennett, Qiang Yang, Brian R Berquist, John P Giddens, Zhongjie Ren, Vally Kommineni, Ryan P Murray, Earl L White, Barry R Holtz, Lai-Xi Wang, Sylvain Marcel
N -glycosylation profoundly affects the biological stability and function of therapeutic proteins, which explains the recent interest in glycoengineering technologies as methods to develop biobetter therapeutics. In current manufacturing processes, N -glycosylation is host-specific and remains difficult to control in a production environment that changes with scale and production batches leading to glycosylation heterogeneity and inconsistency. On the other hand, in vitro chemoenzymatic glycan remodeling has been successful in producing homogeneous pre-defined protein glycoforms, but needs to be combined with a cost-effective and scalable production method...
January 31, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29102080/prospects-of-pasylation%C3%A2-for-the-design-of-protein-and-peptide-therapeutics-with-extended-half-life-and-enhanced-action
#4
Michaela Gebauer, Arne Skerra
Pharmacokinetic (PK) extension is no longer just a means to create improved second generation biologics (so-called biobetters), but constitutes an accepted strategy in biopharmaceutical drug development today. Although PEGylation has become a widely applied methodology to furnish therapeutic proteins and peptides with prolonged plasma half-life, the immunogenicity and missing biodegradability of this synthetic polymer has prompted an evident need for alternatives. PASylation is based on biological polypeptides made of the small l-amino acids Pro, Ala and/or Ser (PAS), which adopt a random coil structure in aqueous buffers with surprisingly similar biophysical properties as PEG...
September 15, 2017: Bioorganic & Medicinal Chemistry
https://www.readbyqxmd.com/read/28972297/progress-in-biopharmaceutical-development
#5
REVIEW
Malgorzata Kesik-Brodacka
Since its introduction in 1982, biopharmaceutical drugs have revolutionized the treatment of a broad spectrum of diseases and are increasingly used in nearly all branches of medicine. In recent years, the biopharmaceuticals market has developed much faster than the market for all drugs and is believed to have great potential for further dynamic growth because of the tremendous demand for these drugs. Biobetters, which contain altered active pharmaceutical ingredients with enhanced efficacy, will play an important role in the development of biopharmaceuticals...
October 3, 2017: Biotechnology and Applied Biochemistry
https://www.readbyqxmd.com/read/28921436/strategies-to-develop-therapeutic-n-and-o-hyperglycosylated-proteins
#6
Agustina Gugliotta, Natalia Ceaglio, Marina Etcheverrigaray, Ricardo Kratje, Marcos Oggero
Glycoengineering by N- and/or O-hyperglycosylation represents a procedure to introduce potential sites for adding N- and/or O-glycosyl structures to proteins with the aim of producing biotherapeutics with improved pharmacodynamic and pharmacokinetic properties. In this chapter, a detailed description of the steps routinely performed to generate new proteins having high content of N- and/or O-glycosyl moieties is carried out. The rational strategy involves the initial stage of designing N- and/or O-hyperglycosylated muteins to be expressed by mammalian cells and includes the upstream and downstream processing stages necessary to develop hyperglycosylated versions of the proteins of interest with the purpose of beginning the long road toward producing biobetters...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28895717/backbone-circularization-coupled-with-optimization-of-connecting-segment-in-effectively-improving-the-stability-of-granulocyte-colony-stimulating-factor
#7
Takamitsu Miyafusa, Risa Shibuya, Wataru Nishima, Rie Ohara, Chuya Yoshida, Shinya Honda
Backbone circularization of protein is a powerful method to improve its structural stability. In this paper, we presumed that a tight connection leads to much higher stability. Therefore, we designed circularized variants of a granulocyte-colony stimulating factor (G-CSF) with a structurally optimized terminal connection. To estimate the appropriate length of the connection, we surveyed the Protein Data Bank to find local structures as a model for the connecting segment. We set the library of local structures composed of "helix-loop-helix," subsequently selected entries similar to the G-CSF terminus, and finally sorted the hit structures according to the loop length...
October 20, 2017: ACS Chemical Biology
https://www.readbyqxmd.com/read/28895363/-characterization-of-n-glycosylation-in-an-anti-egfr-monoclonal-antibody-produced-by-different-expression-systems
#8
Chong Wang, Huaizu Guo
The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. The present study was designed to explore the impact of a different expression system on mAb N-glycosylation. The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS)...
June 25, 2017: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
https://www.readbyqxmd.com/read/28595518/humanization-of-chimeric-anti-cd20-antibody-by-logical-and-bioinformatics-approach-with-retention-of-biological-activity
#9
Yoke L Khoo, Swee H Cheah, Heilly Chong
AIM: To develop a fully bioactive humanized antibody from the chimeric rituximab for potential clinical applications using a relatively simpler and faster logical and bioinformatics approach. METHODS: From bioinformatics data, mismatched mouse amino acids in variable light and heavy chain amphipathic regions were identified and substituted with those common to human antibody framework. Appropriate synthetic DNA sequences inserted into vectors were transfected into HEK293 cells to produce the humanized antibody...
June 2017: Immunotherapy
https://www.readbyqxmd.com/read/28179976/biobetters-from-an-integrated-computational-experimental-approach
#10
REVIEW
Serdar Kuyucak, Veysel Kayser
Biobetters are new drugs designed from existing peptide or protein-based therapeutics by improving their properties such as affinity and selectivity for the target epitope, and stability against degradation. Computational methods can play a key role in such design problems-by predicting the changes that are most likely to succeed, they can drastically reduce the number of experiments to be performed. Here we discuss the computational and experimental methods commonly used in drug design problems, focusing on the inverse relationship between the two, namely, the more accurate the computational predictions means the less experimental effort is needed for testing...
2017: Computational and Structural Biotechnology Journal
https://www.readbyqxmd.com/read/28029136/application-of-lectin-array-technology-for-biobetter-characterization-its-correlation-with-fc%C3%AE-riii-binding-and-adcc
#11
Markus Roucka, Klaus Zimmermann, Markus Fido, Andreas Nechansky
Lectin microarray technology was applied to compare the glycosylation pattern of the monoclonal antibody MB311 expressed in SP2.0 cells to an antibody-dependent cellular cytotoxic effector function (ADCC)-optimized variant (MB314). MB314 was generated by a plant expression system that uses genetically modified moss protoplasts (Physcomitrella patens) to generate a de-fucosylated version of MB311. In contrast to MB311, no or very low interactions of MB314 with lectins Aspergillus oryzae l-fucose (AOL), Pisum sativum agglutinin (PSA), Lens culinaris agglutinin (LCA), and Aleuria aurantia lectin (AAL) were observed...
December 24, 2016: Microarrays
https://www.readbyqxmd.com/read/27916504/the-state-of-play-and-future-of-antibody-therapeutics
#12
REVIEW
Zehra Elgundi, Mouhamad Reslan, Esteban Cruz, Vicki Sifniotis, Veysel Kayser
It has been over four decades since the development of monoclonal antibodies (mAbs) using a hybridoma cell line was first reported. Since then more than thirty therapeutic antibodies have been marketed, mostly as oncology, autoimmune and inflammatory therapeutics. While antibodies are very efficient, their cost-effectiveness has always been discussed owing to their high costs, accumulating to more than one billion dollars from preclinical development through to market approval. Because of this, therapeutic antibodies are inaccessible to some patients in both developed and developing countries...
December 1, 2017: Advanced Drug Delivery Reviews
https://www.readbyqxmd.com/read/26964748/analytical-detection-and-characterization-of-biopharmaceutical-glycosylation-by-ms
#13
REVIEW
Myung Jin Oh, Serenus Hua, Unyong Kim, Hyun Joong Kim, Jua Lee, Jae-Han Kim, Hyun Joo An
Glycosylation plays an important role in ensuring the proper structure and function of most biotherapeutic proteins. Even small changes in glycan composition, structure, or location can have a drastic impact on drug safety and efficacy. Recently, glycosylation has become the subject of increased focus as biopharmaceutical companies rush to create not only biosimilars, but also biobetters based on existing biotherapeutic proteins. Against this backdrop of ongoing biopharmaceutical innovation, updated methods for accurate and detailed analysis of protein glycosylation are critical for biopharmaceutical companies and government regulatory agencies alike...
April 2016: Bioanalysis
https://www.readbyqxmd.com/read/26554886/update-on-biosimilars-of-granulocyte-colony-stimulating-factor-when-no-news-is-good-news
#14
REVIEW
Miriam Schulz, Halvard Bonig
PURPOSE OF REVIEW: With the approval of the first biosimilar granulocyte colony-stimulating factor (G-CSF), biosimilars - copies of therapeutic biologicals whose patent protection has expired - have finally reached the US healthcare market. Its advent is an occasion for a closer look at recent insights into biosimilar G-CSF and an attempt at prognosticating the future (future role) of biosimilars in general. RECENT FINDINGS: Recent literature regarding biosimilar G-CSF orbits significantly around patient access and effects on healthcare expenditure...
January 2016: Current Opinion in Hematology
https://www.readbyqxmd.com/read/26387511/commercial-scale-biotherapeutics-manufacturing-facility-for-plant-made-pharmaceuticals
#15
REVIEW
Barry R Holtz, Brian R Berquist, Lindsay D Bennett, Vally J M Kommineni, Ranjith K Munigunti, Earl L White, Don C Wilkerson, Kah-Yat I Wong, Lan H Ly, Sylvain Marcel
Rapid, large-scale manufacture of medical countermeasures can be uniquely met by the plant-made-pharmaceutical platform technology. As a participant in the Defense Advanced Research Projects Agency (DARPA) Blue Angel project, the Caliber Biotherapeutics facility was designed, constructed, commissioned and released a therapeutic target (H1N1 influenza subunit vaccine) in <18 months from groundbreaking. As of 2015, this facility was one of the world's largest plant-based manufacturing facilities, with the capacity to process over 3500 kg of plant biomass per week in an automated multilevel growing environment using proprietary LED lighting...
October 2015: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/26375407/key-strategic-factors-for-stakeholders-in-the-current-global-biosimilar-market
#16
Daniel Casey
What are the strategies of success in the global biosimilar market? In general, originators have short-term options available, such as patent litigation, but should focus on sustainable long-term strategies such as development of biobetters or incorporating biosimilars as complements to existing products. For new entrants to the market, the learning curve will be steep and only biosimilars developed to compete with the best-selling biologics will be successful over the next decade. The attitudes of physicians, patients and payers will be crucial to the reception future biosimilars will receive and it will be essential that other players contribute to this debate...
February 2016: Drug Discovery Today
https://www.readbyqxmd.com/read/26215462/basal-buffer-systems-for-a-newly-glycosylated-recombinant-human-interferon-%C3%AE-with-biophysical-stability-and-doe-approaches
#17
Nam Ah Kim, Kyoung Song, Dae Gon Lim, Shavron Hada, Young Kee Shin, Sangmun Shin, Seong Hoon Jeong
The purpose of this study was to develop a basal buffer system for a biobetter version of recombinant human interferon-β 1a (rhIFN-β 1a), termed R27T, to optimize its biophysical stability. The protein was pre-screened in solution as a function of pH (2-11) using differential scanning calorimetry (DSC) and dynamic light scattering (DLS). According to the result, its experimental pI and optimal pH range were 5.8 and 3.6-4.4, respectively. Design of experiment (DoE) approach was developed as a practical tool to aid formulation studies as a function of pH (2...
October 12, 2015: European Journal of Pharmaceutical Sciences
https://www.readbyqxmd.com/read/26177629/fusion-proteins-for-half-life-extension-of-biologics-as-a-strategy-to-make-biobetters
#18
REVIEW
William R Strohl
The purpose of making a "biobetter" biologic is to improve on the salient characteristics of a known biologic for which there is, minimally, clinical proof of concept or, maximally, marketed product data. There already are several examples in which second-generation or biobetter biologics have been generated by improving the pharmacokinetic properties of an innovative drug, including Neulasta(®) [a PEGylated, longer-half-life version of Neupogen(®) (filgrastim)] and Aranesp(®) [a longer-half-life version of Epogen(®) (epoetin-α)]...
August 2015: BioDrugs: Clinical Immunotherapeutics, Biopharmaceuticals and Gene Therapy
https://www.readbyqxmd.com/read/26096711/rational-design-of-biobetters-with-enhanced-stability
#19
Fabienne Courtois, Curtiss P Schneider, Neeraj J Agrawal, Bernhardt L Trout
Biotherapeutics are the fastest growing class of pharmaceutical with a rapidly evolving market facing the rise of biosimilar and biobetter products. In contrast to a biosimilar, which is derived from the same gene sequence as the innovator product, a biobetter has enhanced properties, such as enhanced efficacy or reduced immunogenicity. Little work has been carried out so far to increase the intrinsic stability of biotherapeutics via sequence changes, even though, aggregation, the primary degradation pathway of proteins, leads to issues ranging from manufacturing failure to immunological response and to loss of therapeutic activity...
August 2015: Journal of Pharmaceutical Sciences
https://www.readbyqxmd.com/read/26011014/moss-made-pharmaceuticals-from-bench-to-bedside
#20
REVIEW
Ralf Reski, Juliana Parsons, Eva L Decker
Over the past two decades, the moss Physcomitrella patens has been developed from scratch to a model species in basic research and in biotechnology. A fully sequenced genome, outstanding possibilities for precise genome-engineering via homologous recombination (knockout moss), a certified GMP production in moss bioreactors, successful upscaling to 500 L wave reactors, excellent homogeneity of protein glycosylation, remarkable batch-to-batch stability and a safe cryopreservation for master cell banking are some of the key features of the moss system...
October 2015: Plant Biotechnology Journal
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