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K1, mAb

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https://www.readbyqxmd.com/read/27795303/antibody-based-immunotherapy-to-treat-and-prevent-infection-with-hypervirulent-klebsiella-pneumoniae
#1
Elizabeth Diago-Navarro, Isabel Calatayud-Baselga, Donglei Sun, Camille Khairallah, Inderjit Mann, Amaia Ulacia-Hernando, Brian Sheridan, Meiqing Shi, Bettina C Fries
Hypervirulent Klebsiella pneumoniae (hvKp) strains are predicted to become a major threat in Asia if antibiotic resistance continues to spread. Anticapsular antibodies (Abs) were developed because disseminated infections caused by hvKp are associated with significant morbidity and mortality even with antibiotic sensitive strains. K1-serotype polysaccharide capsules (K1-CPS) are expressed by the majority of hvKp strains. In this study K1-CPS specific IgG Abs were generated by conjugation of K1-CPS to immunogenic Anthrax Protective Antigen (PA) protein...
October 26, 2016: Clinical and Vaccine Immunology: CVI
https://www.readbyqxmd.com/read/27704181/reduction-of-ammonia-and-lactate-through-the-coupling-of-glutamine-synthetase-selection-and-downregulation-of-lactate-dehydrogenase-a-in-cho-cells
#2
Soo Min Noh, Jin Hyoung Park, Myung Sin Lim, Jong Won Kim, Gyun Min Lee
Chinese hamster ovary (CHO) cell cultivation for production of therapeutic proteins is accompanied by production of metabolic wastes, mostly ammonia and lactate. To reduce ammonia production, the glutamine synthetase (GS) system was used to develop therapeutic monoclonal antibody (mAb)-producing CHO cells (SM-0.025). Additionally, the lactate dehydrogenase-A (LDH-A) was downregulated with shRNA to reduce lactate production in SM-0.025. The resulting mAb-producing cell lines (#2, #46, and #52) produced less ammonia than the host cell line during the exponential phase due to GS protein overexpression...
October 4, 2016: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/27059141/development-of-a-monoclonal-antibody-specific-to-envelope-domain-iii-with-broad-spectrum-detection-of-all-four-dengue-virus-serotypes
#3
Sae-Hae Kim, Yu Na Kim, Thang Thua Truong, Nguyen Thi Thu Thuy, Le Quynh Mai, Yong-Suk Jang
Dengue virus (DENV) is a mosquito-borne pathogen that annually infects more than 390 million people in 100 different countries. Symptoms of the viral infection include a relatively weak dengue fever to severe dengue hemorrhagic fever/dengue shock syndrome, which are mortal infectious diseases. As of yet, there is no commercially available vaccine or therapeutic for DENV. Currently, passive immunotherapy using DENV-specific antibody (Ab) is a considered strategy to treat DENV infection. Here, we developed a monoclonal Ab (mAb), EDIIImAb-61, specific to the DENV domain III of the envelope glycoprotein (EDIII) with broad-spectrum detection ability to all four DENV serotypes (DENV-1∼4) to use as a therapeutic Ab...
May 13, 2016: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/26361260/mechanisms-of-action-of-tshr-autoantibodies
#4
REVIEW
J Furmaniak, J Sanders, R Núñez Miguel, B Rees Smith
The availability of human monoclonal antibodies (MAbs) to the TSHR has enabled major advances in our understanding of how TSHR autoantibodies interact with the receptor. These advances include determination of the crystal structures of the TSHR LRD in complex with a stimulating autoantibody (M22) and with a blocking type autoantibody (K1-70). The high affinity of MAbs for the TSHR makes them particularly suitable for use as ligands in assays for patient serum TSHR autoantibodies. Also, M22 and K1-70 are effective at low concentrations in vivo as TSHR agonists and antagonists respectively...
September 2015: Hormone and Metabolic Research, Hormon- und Stoffwechselforschung, Hormones et Métabolisme
https://www.readbyqxmd.com/read/26000138/blocking-type-tsh-receptor-antibodies
#5
REVIEW
Jadwiga Furmaniak, Jane Sanders, Bernard Rees Smith
TSH receptor (TSHR) autoantibodies (TRAbs) play a key role in the pathogenesis of Graves' disease. In the majority of patients, TRAbs stimulate thyroid hormone synthesis via activation of the TSHR (stimulating TRAbs, TSHR agonists). In some patients, TRAbs bind to the receptor but do not cause activation (blocking TRAbs, TSHR antagonists). Isolation of human TSHR monoclonal antibodies (MAbs) with either stimulating (M22 and K1-18) or blocking activities (5C9 and K1-70) has been a major advance in studies on the TSHR...
April 2013: Auto- Immunity Highlights
https://www.readbyqxmd.com/read/25658886/recombinant-immunoglobulin-a-specific-for-influenza-a-virus-hemagglutinin-production-functional-analysis-and-formation-of-secretory-immunoglobulin-a
#6
Kentaro Shoji, Tadanobu Takahashi, Kohta Kurohane, Koki Iwata, Takeshi Matsuoka, Shogo Tsuruta, Takatomo Sugino, Masaki Miyake, Takashi Suzuki, Yasuyuki Imai
Secretory immunoglobulin (Ig) A (SIgA), comprised of dimeric IgA and secretory component (SC), is believed to provide a defense mechanism on the mucosal surface. Influenza A virus (IAV) hemagglutinin (HA)-specific SIgA is thought to play an important role in the prevention of IAV infection. However, the topical application of preformed IAV-specific SIgA has not been shown to prevent IAV infection. This is due to the difficulty in the production of antigen-specific IgA monoclonal antibodies (mAbs) and monoclonal SIgA...
April 2015: Viral Immunology
https://www.readbyqxmd.com/read/25651860/fast-track-antibody-v-gene-rescue-recombinant-expression-in-plants-and-characterization-of-a-pfmsp4-specific-antibody
#7
Stephanie Kapelski, Alexander Boes, Holger Spiegel, Melanie de Almeida, Torsten Klockenbring, Andreas Reimann, Rainer Fischer, Stefan Barth, Rolf Fendel
BACKGROUND: Monoclonal antibodies (mAbs) are essential tools in biological research, diagnosis and therapy, and are conventionally produced in murine hybridoma cell lines. Professional applications of mAbs depend on the steady supply of material. Because hybridoma cultures can stop producing the antibody or even die, preservation of the unique epitope specificity of mAbs by rescue of the sequences encoding the antibody variable domains (V regions) is important. The availability of these sequences enables not only the recombinant expression of the original antibody for further applications, but opens the road for antibody engineering towards innovative diagnostic or therapeutic applications...
2015: Malaria Journal
https://www.readbyqxmd.com/read/25064149/calcium-sensitive-immunoaffinity-chromatography-gentle-and-highly-specific-retrieval-of-a-scarce-plasma-antigen-collectin-lk-cl-lk
#8
Maiken L Henriksen, Kirstine L Madsen, Karsten Skjoedt, Soren Hansen
Immunoaffinity chromatography is a powerful fractionation technique that has become indispensable for protein purification and characterization. However, it is difficult to retrieve bound proteins without using harsh or denaturing elution conditions, and the purification of scarce antigens to homogeneity may be impossible due to contamination with abundant antigens. In this study, we purified the scarce, complement-associated plasma protein complex, collectin LK (CL-LK, complex of collectin liver 1 and kidney 1), by immunoaffinity chromatography using a calcium-sensitive anti-collectin-kidney-1 mAb...
November 2014: Journal of Immunological Methods
https://www.readbyqxmd.com/read/25044602/humanized-antibody-neutralizing-2009-pandemic-h1n1-virus
#9
Nachiket Shembekar, Vamsee V Aditya Mallajosyula, Piyush Chaudhary, Vaibhav Upadhyay, Raghavan Varadarajan, Satish Kumar Gupta
The 2009 pandemic H1N1 S-OIV (swine origin influenza A virus) caused noticeable morbidity and mortality worldwide. In addition to vaccine and antiviral drug therapy, the use of influenza virus neutralizing monoclonal antibodies (MAbs) for treatment purposes is a viable alternative. We previously reported the isolation of a high affinity, potently neutralizing murine MAb MA2077 against 2009 pandemic H1N1 virus. We describe here the humanization of MA2077 and its expression in a mammalian cell line. Six complementarity-determining regions (CDRs) of MA2077 were grafted onto the human germline variable regions; along with six and eight back mutations in the framework of heavy and light chains, respectively, pertaining to the vernier zone and interchain packing residues to promote favorable CDR conformation and facilitate antigen binding...
December 2014: Biotechnology Journal
https://www.readbyqxmd.com/read/24603121/characterization-of-escherichia-coli-k1-colominic-acid-specific-murine-antibodies-that-are-cross-protective-against-neisseria-meningitidis-groups-b-c-and-y
#10
In Ho Park, Jisheng Lin, Ji Eun Choi, Jeon-Soo Shin
The capsular polysaccharide (PS) of Neisseria meningitidis serogroup B (NMGB) is α(2-8)-linked N-acetylneuraminic acid (Neu5Ac), which is almost identical to the O-acetylated colominic acid (CA) of Escherichia coli K1 Although E. coli K1 has long been known to elicit cross-protective antibodies against NMGB, limited information on these highly cross-reactive antibodies is available. In the present study, six new monoclonal antibodies (mAbs) specific to both E. coli K1 CA and NMGB PS were produced by immunizing Balb/c mice with E...
June 2014: Molecular Immunology
https://www.readbyqxmd.com/read/23711520/evaluation-of-macroporous-and-microporous-carriers-for-cho-k1-cell-growth-and-monoclonal-antibody-production
#11
Maria Elisa Rodrigues, Ana Rita Costa, Pedro Fernandes, Mariana Henriques, Philip Cunnah, David W Melton, Joana Azeredo, Rosário Oliveira
The emergence of microcarrier technology has brought a renewed interest in anchorage-dependent cell culture for high-yield processes. Well-known in vaccine production, microcarrier culture also has potential for application in other fields. In this work, two types of microcarriers were evaluated for small-scale monoclonal antibody (mAb) production by CHOK1 cells. Cultures (5 ml) of microporous Cytodex 3 and macroporous CultiSpher-S carriers were performed in vented conical tubes and subsequently scaled-up (20 ml) to shake-flasks, testing combinations of different culture conditions (cell concentration, microcarrier concentration, rocking methodology, rocking speed, and initial culture volume)...
September 28, 2013: Journal of Microbiology and Biotechnology
https://www.readbyqxmd.com/read/23651948/transcriptomic-analysis-of-clonal-growth-rate-variation-during-cho-cell-line-development
#12
Padraig Doolan, Colin Clarke, Paula Kinsella, Laura Breen, Paula Meleady, Mark Leonard, Lin Zhang, Martin Clynes, Sinead T Aherne, Niall Barron
The selection of clones displaying a high rate of cell growth is an essential component of Chinese hamster ovary (CHO) cell line development. In recent years various "omics" technologies have been utilised to understand the mechanisms underlying bioprocess phenotypes. In this study, gene expression analysis using a CHO-specific microarray was conducted for a panel of CHO-K1 MAb-secreting cell lines spanning a range of growth rates that were derived from a single cell line development project. In-silico functional analysis of the resulting transcriptomic data revealed the overrepresentation of biological processes such as cell cycle and translation within those genes upregulated during fast growth, while genes associated with cellular homeostasis were downregulated...
July 10, 2013: Journal of Biotechnology
https://www.readbyqxmd.com/read/23606666/chinese-hamster-ovary-k1-host-cell-enables-stable-cell-line-development-for-antibody-molecules-which-are-difficult-to-express-in-duxb11-derived-dihydrofolate-reductase-deficient-host-cell
#13
Zhilan Hu, Donglin Guo, Shirley S M Yip, Dejin Zhan, Shahram Misaghi, John C Joly, Bradley R Snedecor, Amy Y Shen
Therapeutic monoclonal antibodies (mAb) are often produced in Chinese hamster ovary (CHO) cells. Three commonly used CHO host cells for generating stable cell lines to produce therapeutic proteins are dihydrofolate reductase (DHFR) positive CHOK1, DHFR-deficient DG44, and DUXB11-based DHFR deficient CHO. Current Genentech commercial full-length antibody products have all been produced in the DUXB11-derived DHFR-deficient CHO host. However, it has been challenging to develop stable cell lines producing an appreciable amount of antibody proteins in the DUXB11-derived DHFR-deficient CHO host for some antibody molecules and the CHOK1 host has been explored as an alternative approach...
July 2013: Biotechnology Progress
https://www.readbyqxmd.com/read/23509073/chinese-hamster-ovary-cho-k1-host-cell-enables-stable-cell-line-development-for-antibody-molecules-which-are-difficult-to-express-in-duxb11-derived-dihydrofolate-reductase-dhfr-deficient-host-cell
#14
Zhilan Hu, Donglin Guo, Shirley S M Yip, Dejin Zhan, Shahram Misaghi, John C Joly, Bradley R Snedecor, Amy Y Shen
Therapeutic monoclonal antibodies (mAb) are often produced in Chinese hamster ovary (CHO) cells. Three commonly used CHO host cells for generating stable cell lines to produce therapeutic proteins are dihydrofolate reductase (DHFR) positive CHOK1, DHFR-deficient DG44, and DUXB11-based DHFR deficient CHO. Current Genentech commercial full-length antibody products have all been produced in the DUXB11-derived DHFR-deficient CHO host. However, it has been challenging to develop stable cell lines producing an appreciable amount of antibody proteins in the DUXB11-derived DHFR-deficient CHO host for some antibody molecules and the CHOK1 host has been explored as an alternative approach...
March 19, 2013: Biotechnology Progress
https://www.readbyqxmd.com/read/23487430/the-impact-of-microcarrier-culture-optimization-on-the-glycosylation-profile-of-a-monoclonal-antibody
#15
Ana Rita Costa, Joanne Withers, Maria Elisa Rodrigues, Niaobh McLoughlin, Mariana Henriques, Rosário Oliveira, Pauline M Rudd, Joana Azeredo
Microcarriers are widely used for the large-scale culture of attachment-dependent cells with increased cell densities and, ultimately, higher product yield. In these processes, the specific culture conditions can affect the quality of the product, which is closely related to its glycosylation pattern. Furthermore, the lack of studies in the area reinforces the need to better understand the effects of microcarrier culture in product glycosylation. Consequently, in this work, the glycosylation profile of a monoclonal antibody (mAb) produced by adherent CHO-K1 cells grown in Cytodex 3 was evaluated under different conditions, and compared to that obtained of typical adherent cultures...
December 2013: SpringerPlus
https://www.readbyqxmd.com/read/23478323/resistance-of-the-brain-to-escherichia-coli-k1-infection-depends-on-myd88-signaling-and-the-contribution-of-neutrophils-and-monocytes
#16
Sandra Ribes, Tommy Regen, Tanja Meister, Simone C Tauber, Sandra Schütze, Alexander Mildner, Matthias Mack, Uwe-Karsten Hanisch, Roland Nau
Escherichia coli is the leading cause of Gram-negative neonatal bacterial meningitis and also causes meningitis and meningoencephalitis in older and immunocompromised patients. Here, we determined the contribution of granulocytes, monocytes, and TLR signaling cascades in the resistance of adult mice to Escherichia coli K1 brain infection. Deficiency in MyD88 (myd88(-/-)) but not in TRIF (trif(lps2)) adaptor proteins dramatically reduced the survival of animals. Depletion of CD11b(+) Ly-6G(+) Ly-6C(int) neutrophils by application of the anti-Ly-6G (1A8) monoclonal antibody (MAb) led to higher bacterial loads in cerebellum and spleen tissue and resulted in increased mortality compared to those of isotype-treated controls...
May 2013: Infection and Immunity
https://www.readbyqxmd.com/read/23306891/advances-and-drawbacks-of-the-adaptation-to-serum-free-culture-of-cho-k1-cells-for-monoclonal-antibody-production
#17
Maria Elisa Rodrigues, Ana Rita Costa, Mariana Henriques, Philip Cunnah, David W Melton, Joana Azeredo, Rosário Oliveira
Currently, mammalian cell technology has become the focus of biopharmaceutical production, with strict regulatory scrutiny of the techniques employed. Major concerns about the presence of animal-derived components in the culture media led to the development of serum-free (SF) culture processes. However, cell adaptation to SF conditions is still a major challenge and limiting step of process development. Thus, this study aims to assess the impact of SF adaptation on monoclonal antibody (mAb) production, identify the most critical steps of cell adaptation to the SF EX-CELL medium, and create basic process guidelines...
February 2013: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/23297385/the-k1-capsular-polysaccharide-from-acinetobacter-baumannii-is-a-potential-therapeutic-target-via-passive-immunization
#18
Thomas A Russo, Janet M Beanan, Ruth Olson, Ulrike MacDonald, Andrew D Cox, Frank St Michael, Evgeny V Vinogradov, Brad Spellberg, Nicole R Luke-Marshall, Anthony A Campagnari
The emergence of extremely resistant and panresistant Gram-negative bacilli, such as Acinetobacter baumannii, requires consideration of nonantimicrobial therapeutic approaches. The goal of this report was to evaluate the K1 capsular polysaccharide from A. baumannii as a passive immunization target. Its structure was determined by a combination of mass spectrometric and nuclear magnetic resonance (NMR) techniques. Molecular mimics that might raise the concern for autoimmune disease were not identified. Immunization of CD1 mice demonstrated that the K1 capsule is immunogenic...
March 2013: Infection and Immunity
https://www.readbyqxmd.com/read/23247406/the-impact-of-cell-adaptation-to-serum-free-conditions-on-the-glycosylation-profile-of-a-monoclonal-antibody-produced-by-chinese-hamster-ovary-cells
#19
Ana Rita Costa, Joanne Withers, Maria Elisa Rodrigues, Niaobh McLoughlin, Mariana Henriques, Rosário Oliveira, Pauline M Rudd, Joana Azeredo
N-glycosylation is one of the most crucial parameters affecting the biological activity of therapeutic monoclonal antibodies (mAbs), and should therefore be closely monitored and controlled to guarantee a consistent and high-quality product in biopharmaceutical processes. In the present work, the effect of the time-consuming step of gradual cell adaptation to serum-free conditions on the glycosylation profile of a mAb produced by CHO-K1 cells was evaluated. High-performance liquid chromatography analysis revealed important changes in mAb glycosylation patterns in all steps of serum reduction...
June 25, 2013: New Biotechnology
https://www.readbyqxmd.com/read/22902388/comparison-of-commercial-serum-free-media-for-cho-k1-cell-growth-and-monoclonal-antibody-production
#20
COMPARATIVE STUDY
Maria Elisa Rodrigues, Ana Rita Costa, Mariana Henriques, Joana Azeredo, Rosário Oliveira
The selection of a serum-free medium for a particular process of production using mammalian cells is a critical step for its success. In this study, seven commercially available serum-free media (EX-CELL, ISF-I, CD CHO, CDM4CHO, CHO-III-A, Octomed and HybridoMed) were evaluated and compared for cell growth and monoclonal antibody (mAb) production of a transfected CHO-K1 cell line. In the conditions assayed, EX-CELL and particularly CDM4CHO are the most recommended media for extended biopharmaceutical processes, on account of inducing superior levels of cell proliferation and mAb production, accentuated by a tendency to improve over time...
November 1, 2012: International Journal of Pharmaceutics
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