Characterization of recombinant human diamine oxidase (rhDAO) produced in Chinese Hamster Ovary (CHO) cells

Gludovacz, Elisabeth; Maresch, Daniel; Bonta, Maximilian; Szöllösi, Helen; Furtmüller, Paul G.; McCarley, Robert W.; Altmann, Friedrich; Limbeck, Andreas; Borth, Nicole; Jilma, Bernd; Boehm, Thomas

doi:10.1016/j.jbiotec.2016.04.002

Cited by 20 publications

(26 citation statements)

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“…DAO activity was measured using a chemiluminescence‐based enzyme activity assay 14 as described 50 . Briefly, DAO released hydrogen peroxide is used to oxidize luminol by horseradish peroxidase.…”

Section: Methodsmentioning

confidence: 99%

“…Bound human DAO was detected with a polyclonal rabbit IgG serum fraction diluted 1 to 1000. Rabbits were immunized with purified recombinant human DAO 50 and serum IgGs isolated using standard procedures (Eurogentec Polyclonal Antibody Production Service, Seraing, Belgium). The bound rabbit antibodies were detected with a 1 to 32000 dilution of donkey anti-rabbit IgG HRP-labelled antibodies (SAB3700928, Sigma-Aldrich, Austria) in the presence of 10 µg/ml donkey IgG (ChromPure #017-000-003; Jackson ImmunoResearch).…”

Section: Methodsmentioning

confidence: 99%

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Pregnancy-associated diamine oxidase originates from extravillous trophoblasts and is decreased in early-onset preeclampsia

Velicky

Windsperger

Petroczi

et al. 2018

Sci Rep

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Human extravillous trophoblast (EVT) invasion of the pregnant uterus constitutes a pivotal event for the establishment of the maternal-fetal interface. Compromised EVT function manifesting in inadequate arterial remodeling is associated with the severe pregnancy disorder early-onset preeclampsia (eoPE). Recent studies suggest that EVTs invade the entire uterine vasculature including arteries, veins and lymphatics in the first trimester of pregnancy. We therefore hypothesized that EVT-derived factors accumulate in the circulation of pregnant women early in gestation and may serve to predict eoPE. In contrast to published literature, we demonstrate that placenta-associated diamine oxidase (DAO) is not expressed by maternal decidual cells but solely by EVTs, especially when in close proximity to decidual vessels. Cultures of primary EVTs express and secret large amounts of bioactive DAO. ELISA measurements indicate a pregnancy-specific rise in maternal DAO plasma levels around gestational week (GW) 7 coinciding with vascular invasion of EVTs. Strikingly, DAO levels from eoPE cases were significantly lower (40%) compared to controls in the first trimester of pregnancy but revealed no difference at mid gestation. Furthermore, DAO-containing pregnancy plasma rapidly inactivates pathophysiologically relevant histamine levels. This study represents the first proof of concept suggesting EVT-specific signatures as diagnostic targets for the prediction of eoPE.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Pregnancy-associated diamine oxidase originates from extravillous trophoblasts and is decreased in early-onset preeclampsia

Velicky

Windsperger

Petroczi

et al. 2018

Sci Rep

Self Cite

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“…The subclones were isolated after multiple rounds of cell sorting for increased transient productivity 27. Producer cell lines were CHO‐S‐Humira, producing the human monoclonal antibody Adalimumab and CHO‐K1‐hDAO expressing the copper‐containing human diamine oxidase 28. All cell lines were cultured in CD‐CHO medium (Gibco ® ), supplemented with 8 mM L‐glutamine (Merck Millipore) and 0.2% Anti‐Clumping Agent (Gibco ® ) in shaker flasks at 140 rpm, 37°C, 7% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

Label‐free live cell imaging by Confocal Raman Microscopy identifies CHO host and producer cell lines

Mateu

Harreither

Schosserer

et al. 2016

Biotechnology Journal

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As a possible viable and non‐invasive method to identify high producing cells, Confocal Raman Microscopy was shown to be able to differentiate CHO host cell lines and derivative production clones. Cluster analysis of spectra and their derivatives was able to differentiate between different producer cell lines and a host, and also distinguished between an intracellular region of high lipid and protein content that in structure resembles the Endoplasmic Reticulum. This ability to identify the ER may be a major contributor to the identification of high producers. PCA enabled the discrimination even of host cell lines and their subclones with inherently higher production capacity. The method is thus a promising option that may contribute to early, non‐invasive identification of high potential candidates during cell line development and possibly could also be used for proof of identity of established production clones.

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“…Diamine oxidase concentrations were measured using a custom-assembled ELISA as described recently 14 using recombinant human diamine oxidase as a standard 15 . Briefly, a purified monoclonal antibody raised against human DAO isolated from Caco-2 cell supernatant was coated at 5 µg/ml onto white high protein binding microtiter plates (Greiner bio-one 655074) in 50 mM carbonate-bicarbonate coating buffer pH 9.6 overnight at 4 °C.…”

Section: Sample Analysismentioning

confidence: 99%

Regulation of histamine and diamine oxidase in patients undergoing orthotopic liver transplantation

Schiefer

Baron

Boehm

et al. 2020

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increased concentrations of the vasodilator histamine have been observed in patients undergoing abdominal surgery. the role of histamine during orthotopic liver transplantation (oLt) has only been studied in animals. the aim of this study was to measure plasma concentrations of histamine and its degrading enzyme diamine oxidase (DAo) in patients undergoing orthotopic liver transplantation, and assess whether histamine or DAo correlate with intraoperative noradrenaline requirements. Histamine and DAO concentrations were measured in 22 adults undergoing liver transplantation and 22 healthy adults. Furthermore, norepinephrine requirements during liver transplantation were recorded. Baseline concentrations of histamine and DAo were greater in patients, who underwent liver transplantation, than in healthy individuals (Histamine: 6.4 nM, IQR[2.9-11.7] versus 4.3 nM, IQR[3.7-7.1], p = 0.029; DAO: 2.0 ng/mL, IQR[1.5-4.1] versus <0,5 ng/mL, IQR[<0.5-1.1], p < 0.001). During liver transplantation, histamine concentrations decreased to 1.8 nM, IQR[0.5-4.9] in the anhepatic phase (p < 0.0001 versus baseline), and to 1.5 nM, IQR[0.5-2.9] after reperfusion (p < 0.0001 versus baseline). In contrast, DAO concentrations increased to 35.5 ng/ml, IQR[20-50] in the anhepatic phase (p = 0.001 versus baseline) and to 39.5 ng/ml, IQR[23-64] after reperfusion (p = 0.001 versus baseline), correlating inversely with histamine. norepinephrine requirements during human liver transplantation correlated significantly with DAO concentrations in the anhepatic phase (r = 0.58, p = 0.011) and after reperfusion (r = 0.56; p = 0.022). In patients undergoing orthotopic liver transplantation, histamine concentrations decrease whereas DAo concentrations increase manifold. Diamine oxidase correlates with intraoperative norepinephrine requirements in patients undergoing oLt. Orthotopic liver transplantation (OLT) is the only curative procedure for end-stage liver disease (ESLD). Patients with ESLD usually present with a systemic hyperdynamic cardiovascular regulation, demonstrating an increased cardiac output, reduced systemic vascular resistance and impaired vascular responsiveness to stress. During OLT, patients are exposed to further major hemodynamic alterations primarily due to temporary caval clamping, blood loss and ischemia/reperfusion (I/R) injury. Temporary clamping of the inferior vena cava decreases cardiac output by approximately 50% due to a reduction in venous return to the heart 1,2 , while blood loss leads to hypovolemia. I/R injury, mainly characterized by oxidative damage and a severe inflammatory response, triggers the secretion of vasoactive mediators into the recipients' systemic circulation 3. Thus, OLT is frequently associated with hemodynamic instability of the recipient presenting clinically with hypotension, increased vasopressor support and cardiac arrhythmias.

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Characterization of recombinant human diamine oxidase (rhDAO) produced in Chinese Hamster Ovary (CHO) cells

Cited by 20 publications

References 56 publications

Pregnancy-associated diamine oxidase originates from extravillous trophoblasts and is decreased in early-onset preeclampsia

Pregnancy-associated diamine oxidase originates from extravillous trophoblasts and is decreased in early-onset preeclampsia

Label‐free live cell imaging by Confocal Raman Microscopy identifies CHO host and producer cell lines

Regulation of histamine and diamine oxidase in patients undergoing orthotopic liver transplantation

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