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F T Lim, S M Lim, K Ramasamy
There is growing interest in the use of probiotic lactic acid bacteria (LAB) for prevention of hypercholesterolaemia. This study assessed the cholesterol lowering ability of Pediococcus acidilactici LAB4 and Lactobacillus plantarum LAB12 in growth media. Both LAB yielded >98% (39.2 μg/ml) cholesterol lowering in growth media. Nile Red staining indicated direct assimilation of cholesterol by the LAB. The LAB were then explored for their prophylactic (pre-treatment of HT29 cells with LAB prior to cholesterol exposure) and biotherapeutic (treatment of HT29 cells with LAB after exposure to cholesterol) use against short and prolonged exposure of HT29 cells to cholesterol, respectively...
December 1, 2016: Beneficial Microbes
Namita Rokana, Rajbir Singh, Rashmi Hogarehalli Mallappa, Virender Kumar Batish, Sunita Grover
Probiotic Lactobacillus plantarum MTCC 5690 -a probiotic strain of Indian gut origin as such and milk formulations produced with the same were explored in this sudy as biotherapeutics by evauating their functional efficacy against Salmonella infection in mice. The efficacy of milk formulations (fermented/unfermented) of MTCC 5690 for enhancement of intestinal barrier function was determined by monitoring the permeability and histopathology of intestine. The mice fed with probiotic Dahi (PD), Fermented Probiotic Drink (FD) and Sweetened fermented probiotic drink (SFD) maintained the health and integrity of intestinal epithelium in the infected mice as compared to those fed with phosphate buffered saline, milk, unfermented probiotic milk and Dahi...
October 17, 2016: Journal of Medical Microbiology
Jerrard M Hayes, Mark R Wormald, Pauline M Rudd, Gavin P Davey
Therapeutic antibodies hold great promise for the treatment of cancer and autoimmune diseases, and developments in antibody-drug conjugates and bispecific antibodies continue to enhance treatment options for patients. Immunoglobulin (Ig) G antibodies are proteins with complex modifications, which have a significant impact on their function. The most important of these modifications is glycosylation, the addition of conserved glycans to the antibody Fc region, which is critical for its interaction with the immune system and induction of effector activities such as antibody-dependent cell cytotoxicity, complement activation and phagocytosis...
2016: Journal of Inflammation Research
Jeanette Roman, Silikhone Bouaraphan, Thorsten Verch
AIM: Biologics development often requires multiple immunoassays to evaluate both assay reagents and potential drug candidates resulting in extensive analytical development. METHODOLOGY: We developed a semi-universal, 5-layer platform assay on Gyrolab using secondary antispecies or anti-isotype-specific capture and detection antibodies. We applied the assay to several multivalent vaccines. RESULTS: Method performance exhibited a median accuracy of 110%, reproducibility of 9% CV and intermediate precision of 11% CV...
December 2016: Bioanalysis
Hooman Hefzi, Kok Siong Ang, Michael Hanscho, Aarash Bordbar, David Ruckerbauer, Meiyappan Lakshmanan, Camila A Orellana, Deniz Baycin-Hizal, Yingxiang Huang, Daniel Ley, Veronica S Martinez, Sarantos Kyriakopoulos, Natalia E Jiménez, Daniel C Zielinski, Lake-Ee Quek, Tune Wulff, Johnny Arnsdorf, Shangzhong Li, Jae Seong Lee, Giuseppe Paglia, Nicolas Loira, Philipp N Spahn, Lasse E Pedersen, Jahir M Gutierrez, Zachary A King, Anne Mathilde Lund, Harish Nagarajan, Alex Thomas, Alyaa M Abdel-Haleem, Juergen Zanghellini, Helene F Kildegaard, Bjørn G Voldborg, Ziomara P Gerdtzen, Michael J Betenbaugh, Bernhard O Palsson, Mikael R Andersen, Lars K Nielsen, Nicole Borth, Dong-Yup Lee, Nathan E Lewis
Chinese hamster ovary (CHO) cells dominate biotherapeutic protein production and are widely used in mammalian cell line engineering research. To elucidate metabolic bottlenecks in protein production and to guide cell engineering and bioprocess optimization, we reconstructed the metabolic pathways in CHO and associated them with >1,700 genes in the Cricetulus griseus genome. The genome-scale metabolic model based on this reconstruction, iCHO1766, and cell-line-specific models for CHO-K1, CHO-S, and CHO-DG44 cells provide the biochemical basis of growth and recombinant protein production...
November 23, 2016: Cell Systems
Partha Laskar, Joykrishna Dey, Payel Banik, Mahitosh Mandal, Sudip Kumar Ghosh
Stimuli-sensitive polymeric vesicles or polymersomes as self-assembled colloidal nanocarriers have received paramount importance for their integral role as delivery system for therapeutics and biotherapeutics. This work describes spontaneous polymersome formation at pH 7, as evidenced by surface tension, steady state fluorescence, dynamic light scattering, and microscopic studies, by three hydrophilic random cationic copolymers synthesized using N,N-(dimethylamino)ethyl methacrylate (DMAEM) and methoxy poly(ethylene glycol) monomethacrylate in different mole ratios...
November 23, 2016: Macromolecular Bioscience
Srivalleesha Mallidi, Sriram Anbil, Anne-Laure Bulin, Girgis Obaid, Megumi Ichikawa, Tayyaba Hasan
Photodynamic therapy (PDT) is a photochemistry based treatment modality that involves the generation of cytotoxic species through the interactions of a photosensitizer molecule with light irradiation of an appropriate wavelength. PDT is an approved therapeutic modality for several cancers globally and in several cases has proved to be effective where traditional treatments have failed. The key parameters that determine PDT efficacy are 1. the photosensitizer (nature of the molecules, selectivity, and macroscopic and microscopic localization etc...
2016: Theranostics
Sarah Rogstad, Anneliese Faustino, Ashley Ruth, David Keire, Michael Boyne, Jun Park
The characterization sections of biologics license applications (BLAs) approved by the United States Food and Drug Administration (FDA) between 2000 and 2015 were investigated to examine the extent of the use of mass spectrometry. Mass spectrometry was found to be integral to the characterization of these biotherapeutics. Of the 80 electronically submitted monoclonal antibody and protein biotherapeutic BLAs included in this study, 79 were found to use mass spectrometric workflows for protein or impurity characterization...
November 21, 2016: Journal of the American Society for Mass Spectrometry
Seema C Kumar, Jason A DelCarpini, Qiang Qu, Martin Kane, Boris Gorovits
Biotherapeutics are known for their potential to induce drug specific immune responses, which are commonly evaluated by the detection of anti-drug antibodies (ADAs). For some biotherapeutics, pre-existing ADAs against drug have been observed in drug-naïve matrix. The presence of pre-existing drug specific antibodies may significantly complicate assessment of the screening ADA assay cutpoint value, which is usually established based on the statistical analysis of signal distribution from the drug-naïve individuals...
November 21, 2016: AAPS Journal
Agustina Gugliotta, Natalia Ceaglio, Brenda Raud, Guillermina Forno, Laura Mauro, Ricardo Kratje, Marcos Oggero
Both CHO and HEK cells are interesting hosts for the production of biotherapeutics due to their ability to introduce post-translational modifications such as glycosylation. Even though oligosaccharide structures attached to proteins are conserved among eukaryotes, many differences have been found between therapeutic glycoproteins expressed in hamster and human derived cells. In this work, a hyperglycosylated IFN-α2b mutein (IFN4N) was produced in CHO and HEK cell lines and an extensive characterization of their properties was performed...
November 17, 2016: European Journal of Pharmaceutics and Biopharmaceutics
Kirsty D Ratanji, Jeremy P Derrick, Ian Kimber, Robin Thorpe, Meenu Wadhwa, Rebecca J Dearman
The production of anti-drug antibodies can impact significantly upon the safety and efficacy of biotherapeutics. It is known that various factors, including aggregation and the presence of process-related impurities can modify and augment the immunogenic potential of proteins. The purpose of the investigations reported here was to characterize in mice the influence of aggregation and host cell protein (HCP) impurities on the immunogenicity of a humanized single chain antibody variable fragment (scFv), and mouse albumin...
November 10, 2016: Immunology
Imen Kahouli, Meenakshi Malhotra, Susan Westfall, Moulay A Alaoui-Jamali, Satya Prakash
Probiotics have been shown to have beneficial properties in attenuating the risk of colorectal cancer (CRC) development. However, functional evidence to support such effects for some probiotic bacteria are relatively unknown. Here, we document a significant antioxidant, anti-proliferative and pro-apoptotic activities of Lactobacillus acidophilus ATCC 314 and Lactobacillus fermentum NCIMB 5221 on CRC cells, particularly when used in combination (La-Lf). Furthermore, a superior synergistic activity on the inhibition of tumor growth and modulation of cell proliferation and epithelial markers in the Apc (Min/+) CRC mouse model was explored, based on the expression levels of Ki-67, E-cadherin, β-catenin, and cleaved caspase-3 (CC3) proteins...
November 11, 2016: Applied Microbiology and Biotechnology
Keith O'Brien
MicroRNAs (miRNAs) have gained a lot of interest as biomarkers and biotherapeutics in recent years. The discovery of miRNAs in circulation as recently as 2008, aided by rapid advances in high-throughput profiling techniques initiated an explosion of investigations dedicated to discovering circulating miRNAs as minimally invasive biomarkers. As miRNAs regulate many cellular processes, investigators are actively exploring their relevance in disease treatment by miRNA restoration. This chapter demonstrates an approach to discover miRNAs of biomarker and therapeutic potential by isolating miRNAs from cell lines, performing global miRNA profiling, investigating miRNA expression in clinical specimens and examining their therapeutic relevance by restoring miRNA expression using Lipofectamine in vitro...
2017: Methods in Molecular Biology
Charlotte M Deane, Maximiliano Vásquez
No abstract text is available yet for this article.
November 7, 2016: MAbs
Lynn Conley, Yinying Tao, Alexis Henry, Edward Koepf, Douglas Cecchini, John Pieracci, Sanchayita Ghose
Inclusion of a detergent in protein biotherapeutic purification processes is a simple and very robust method for inactivating enveloped viruses. The detergent Triton X-100 has been used for many years and is part of the production process of several commercial therapeutic proteins. However, recent ecological studies have suggested that Triton X-100 and its break-down products can potentially behave as endocrine disrupters in aquatic organisms, raising concerns from an environmental impact perspective. As such, discharge of Triton X-100 into the waste water treatment plants is regulated in some jurisdictions, and alternative detergents for viral inactivation are required...
November 1, 2016: Biotechnology and Bioengineering
Rong Liu, Brian Hoffpauir, Shannon D Chilewski, Janice Gamberdella, Uma Kavita, Jia Duo, Carol Gleason, Yan Zhang, Renuka Pillutla, Binodh DeSilva, Lora Hamuro
The Gyrolab™ xP is a microfluidic platform for conducting ligand binding assays (LBAs) and is recognized for its utility in discovery bioanalysis. However, few reports have focused on the technology for regulated bioanalysis. This technology has the advantage of low reagent consumption, low sample volume, and automated ligand binding methods. To improve bioanalysis testing timelines and increase the speed at which biotherapeutics are delivered to patients, we evaluated the technology for its potential to deliver high-quality data at reduced testing timelines for regulated bioanalysis...
October 27, 2016: AAPS Journal
J Porter Hunt, Seung Ook Yang, Kristen M Wilding, Bradley C Bundy
Recently reported shelf-stable, on-demand protein synthesis platforms are enabling new possibilities in biotherapeutics, biosensing, biocatalysis, and high throughput protein expression. Lyophilized cell-free protein expression systems not only overcome cold-storage limitations, but also enable stockpiling for on-demand synthesis and completely sterilize the protein synthesis platform. Recently reported high-yield synthesis of cytotoxic protein Onconase from lyophilized E. coli extract preparations demonstrates the utility of lyophilized cell-free protein expression and its potential for creating on-demand biotherapeutics, vaccines, biosensors, biocatalysts, and high throughput protein synthesis...
October 28, 2016: Bioengineered
Frank Loganzo, Matthew Sung, Hans-Peter Gerber
Drug resistance limits the effectiveness of cancer therapies. Despite attempts to develop curative anticancer treatments, tumors evolve evasive mechanisms limiting durable responses. Hence, diverse therapies are used to attack cancer, including cytotoxic and targeted agents. Antibody-drug conjugates (ADC) are biotherapeutics designed to deliver potent cytotoxins to cancer cells via tumor-specific antigens. Little is known about the clinical manifestations of drug resistance to this class of therapy; however, recent preclinical studies reveal potential mechanisms of resistance...
October 25, 2016: Molecular Cancer Therapeutics
Xavier LeSaout, Matteo Costioli, Lynn Jordan, Jeremy Lambert, Ross Beighley, Laurel Provencher, Brian Gerwe, Kevin McGuire, Nico Verlinden, Andrew Barry
Small-scale protein purification presents opportunities for accelerated process development of biotherapeutic molecules. Miniaturization of purification conditions reduces time and allows for parallel processing of samples, thus offering increased statistical significance and greater breadth of variables. The ability of the miniaturized platform to be predictive of larger scale purification schemes is of critical importance. The PerkinElmer JANUS BioTx Pro and Pro-Plus workstations were developed as intuitive, flexible, and automated devices capable of performing parallel small-scale analytical protein purification...
March 2016: Engineering in Life Sciences
Laszlo Hajba, Eszter Csanky, Andras Guttman
Comprehensive carbohydrate analysis of glycoproteins from human biological samples and biotherapeutics are important from diagnostic and therapeutic points of view. This review summarizes the current state-of-the-art liquid phase separation techniques used in N-glycosylation analysis. The different liquid chromatographic techniques and capillary electrophoresis methods are critically discussed in detail. Miniaturization of these methods is also important to increase throughput and decrease analysis time. The sample preparation and labeling methods for asparagine linked oligosaccharides are also addressed...
November 2, 2016: Analytica Chimica Acta
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