• Biomolecules & Therapeutics
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• v.6 no.1
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• pp.50-55
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• 1998

Antigenic potential of a recombinant human erythropoietin (rhEPO) produced by Dong-A charm. Co. Ltd. was examined by active systemic anaphylaxis (ASA) test in guinea pigs, mouse-rat passive cutaneous anaphylaxis (PCA) reaction and passive hemagglutination (PHA) test. In ASA test, rhEPO induced the signs of restlessness, rubbing or licking nose, sneezing and coughing in the animals immunized with rhEPO 1000 lU/kg alone or rhEPO 1000 lU/kg incorporated into Freund\\\\`s complete adjuvant. In the mouse-rat PCA test, only one of six sera from the animals immunized with rhEPO 1000 lUng incorporated into Alum showed positive result. In the PHA test, rhEPO revealed negative results in all of the rhEPO-immunized groups. From these results, rhEPO was considered to produce IgE in guinea pigs and mice, but not IgG and/or IsM in mice. The results of this study were similar to those of the other recombinant human erythropoietin and these positive results were thought to be caused due to the fact that rhEPO were heterogeneous proteins to guinea pigs and mice. Considering the fact that rhErO has an identical structure with indigenous human erythropoietin, rhEPO is not thought to cause immunological problems in clinical use.

• KSBB Journal
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• v.32 no.2
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• pp.90-95
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• 2017

Mammalian cell cultures have been used extensively to produce proteins for therapeutic agent because of their ability to perform post-translational modification including glycosylation. To produce recombinant protein, many factors and parameter are considered such as media composition, host cell type, and culture process. In this study, recombinant human erythropoietin (rhEPO) producing cell line was established by using glutamine synthetase system. To reduce serum concentration in media, we compared direct adaptation with step adaptation. Cell growth was faster in step adaptation. In low-level serum media, there were insufficient glucose for cell growth. Thus, we added glucose in low-level serum media from 2 g/L to 4.5 g/L. Titer of rhEPO was higher than other conditions at 4.5 g/L of glucose. Additionally, N-methyl-D-aspartate (NMDA), 13-cis-retinal, and pluronic F-68 (PF-68) were added to enhance productivity in CHO cell cultures. In conclusion, we applied CHO cell producing rhEPO to low-level of serum in media using step-adaptation. Also, we confirmed positive effect of NMDA, 13-cis-retinal, and PF-68.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.37 no.1
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• pp.9-14
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• 2011

Recently, the population of patients who refuse transfusion has increased for both religious and non-religious reasons, even in life threatening emergency situations. Their refusal has highlighted the need to develop nonblood transfusion surgery techniques to decrease the risk from blood transfusions. A 57-year woman with an ulcerative lesion on the gingiva of the right upper molar area visited the department of oral and maxillofacial surgery in Dankook University Dental Hospital. After a preliminary evaluation, the patient was diagnosed with squamous cell carcinoma. As she refused blood transfusion during surgery for religious reasons, surgery was planned using recombinant human erythropoietin (rHuEPO) without a blood transfusion. The patient underwent a partial maxillectomy, supraomohyoid neck dissection, free radial forearm flap and split thickness skin graft under general anesthesia. rHuEPO and iron were used before and after surgery. The hemoglobin/hematocrit (Hb/Hct) level, iron (Fe) and total iron-binding capacity (TIBC) were assessed. The patient recovered completely without any blood transfusions. rHuEPO is a viable alternative for patients with religious objections to receiving blood transfusions.

• Archives of Plastic Surgery
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• v.33 no.5
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• pp.621-626
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• 2006

Purpose: Erythropoietin is traditionally known to regulate erythropoiesis, but recently its protective effect against ischemia-reperfusion injury has been studied mainly in cardiovascular and neuronal systems. This study was planned to investigate the effects of recombinant human erythropoietin on ischemia-reperfusion injury in rat TRAM flap model. Methods: Superiorly based TRAM flap was elevated and ischemic insult was given for four hours. Thirty minutes before reperfusion, single dose recombinant human Erythropoietin(5000IU/kg) was injected via intraperitoneal route in the treatment group. At 24 hours postoperatively, systemic neutrophil count, tissue myeloperoxidase activity, malonyldialdehyde amount, nitric oxide content, tissue water content and histologic finding of inflammation was evaluated. On 10 days postoperatively, flap survival rate, angiogenesis and change in hematocrit level was evaluated. Results: Tissue nitric oxide level was significantly higher and myeloperoxidase activity was significantly lower in the treatment group 24 hours after reperfusion. Tissue water content was significantly lower in the treatment group. Perivascular neutrophil infiltration and intravascular adhesion was marked in the control group. Mean flap survival after ten days was 69% in the treatment group, and 47% in the control group, demonstrating a significant difference. Neovascularization in the treatment group also outnumbered the control group. No significant hematocrit rise was noted ten days after erythropoietin administration. Conclusion: Recombinant human Erythropoietin improved flap survival in ischemia-reperfusion injured rat TRAM flaps, at least partially owing to suppressed inflammation, increased nitric oxide, and enhanced angiogenesis.

• Biomolecules & Therapeutics
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• v.6 no.3
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• pp.317-327
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• 1998

The subchronic toxicity study of rHuEPO, a newly developed recombinant erythropoietin, was investigated for 13 weeks in Beagle dogs intravenously treated with doses of 100, 500 and 2,500 lU/kg/day. There were no significant changes in body weight, food intake, physical and opthalmic examination, urine analysis, etc. Any toxic response was not observed except for enlarged spleen and extramedullary hematopoiesis. These results indicate that the no-observed adverse effect level (NOAEL) of rHuEPO is 100 lU/kg in Beagle dogs.

• Toxicological Research
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• v.12 no.1
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• pp.101-111
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• 1996

The local irritation studies of DA-3285, recombinant human erytropoietin(rHu-EPO), were carried out in rabbits after the following treatment; single application into the conjunctival sac of the eye, single subcutaneous injection, 7-day repeated subcutaneous injection and 8-day repeated infusion into the ear vein. Also, the local irritancy of DA-3285 leaked around vein was studied in mice by single perivascular injection. The results obtained were as follows. In the result of ocular irritation test, DA-3285 could be considered as a non-irritating material. In single and 7-day repeated subcutaneous irritation test, the irritancy of DA-3285 was not so much different from that of saline. The vascular irritancy of DA3285 by 8-day repeated infusion was negligible and similar to that of saline. And the irritancy of DA3285 by perivascular injection was comparable to that of saline. These results indicate that DA-3285 has no irritating activity when injected through subcutaneous or intravenous route for clinical practice as 3.5% solution.

• Biomolecules & Therapeutics
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• v.4 no.4
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• pp.334-338
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• 1996

Antigenicity of DA-3285, human recombinant erythropoietin which was produced from mammalian cells, was examined in guinea pigs and mice. In active systemic anaphylactic test, mild anaphylactic signs were observed in guinea pigs sensitized subcutaneously with DA-3285 or DA-3285 incorporated with complete Freund's adjuvant(CFA) when challenged with high dose(1000 IU/Kg) of DA-3285. Other groups showed negative responses. In mouse-rat passive cutaneous anaphylaxis test, 20% sera of mice immunized with DA-3285 or DA-3285 mixed with aluminum hydroxide(alum) showed mild positive responses. In the case of indirect haemagglutination reaction(IHA) test, when sheep red blood cells coated with DA-3285 was incubated with mouse serum, all the serum samples were showed negative responses. These results suggest that DA-3285 has a very weak antigenic potential and probably would not induce systemic allergic reactions in clinical uses.

• Biomolecules & Therapeutics
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• v.11 no.2
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• pp.126-132
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• 2003

Efficacy and in vivo bioassay of recombinant human erythropoietin (rh-EPO, DWP413) was investigated. Efficacy studies on erythropoiesis were conducted in normal, cisplatin-induced anemic rats and acute hemorrhage - induced anemic rats. Animals were treated intravenously with DWP413 for 5 days, the changes in the number of red blood cells (RBC), hematocrit value (Hct), hemoglobin concentration (Hb) and reticulocyte were examined. In normal rats, at the doses of 50, 250, and 1250 IU/kg/day, in cisplatin-induced anemic rats, at the doses of 50, 100 and 200 IU/kg/day, RBC, Hb, Hct and reticulocyte were increased dose-depen-dently. And in acute hemorrhage-induced anemic rats, DWP413 (150, 450 and 1350 IU/kg/day) significantly increased RBC, Hb, Hct and reticulocytes. In histopathological findings of kidney, cisplatin alone treated rats expressed severe glomerulus and tubular damage. But in the DWP413 treated rats after cisplatin treatment, these were not remarkable compared to cisplatin alone treated rats. In vivo bioassay, DWP413 had 102.43% of bioactivity compared to erythropoietin BRP(Biological Reference Product, European Directorate for the Quality of Medicines). These results suggest that DWP413 might be useful for the therapy of anemia induce by renal failure and acute blood loss.

• Biomolecules & Therapeutics
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• v.4 no.2
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• pp.154-161
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• 1996

General pharmacological properties of LB-00014, an erythropoietin which was produced by recombinant DNA technique in Biotech Research Institute, LG Chemical Ltd. were examined. The administration of LB-00014 (60, 600, 6000 IU/kg, iv) in mice had no effect in general behavior and central nervous system, and no influences on normal body temperature, writhing syndromes induced by 0.7% acetic acid solution and chemoshock produced by strychnine and pentetrazole solution. LB-00014 (60, 600, 6000 IU/kg, iv) given to anesthetized rabbits showed no effect on blood pressure of carotid artery and respiration rates, and it did not influence the responses produced by injection of acetylcholine or epinephme. The administration of LB-00014 (601, 600, 6000 IU/kg, iv) in rats had no effect on the plasma prothrombin time, activated partial thromboplastin time and hemolytic action. The platelet aggregation induced by collagen in human plasma was not influenced by LB-00014 (10, 100, 1000 lU/kg, iv). It showed no direct effect at 100 and 1000IU/m1 in isolated stomach fundus and uterus of rats and ileum of guinea-pig, and it also had no inhibition of contraction produced by acetylcholine, oxytocin, serotonin and histamine in the above-mentioned preparations. It did not influence gastric secretion, pH and acid output at 6000 IU/kg, iv in rats, but showed a significant increase in intestinal propulsion of mice at 6000 IU/kg, iv. Its administration (60, 600, 6000 lU/kg, iv) caused no effect on urine and electrolyte excretion in rats. These results indicate that LB-00014 does not exsert any of serious pharmacological effects.

• Journal of Yeungnam Medical Science
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• v.11 no.1
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• pp.115-126
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• 1994

The recent availability of recombinant human erythropoietin has opened new perspectives in the management of a variety of anemias. Clinical trials have been initiated in several countries using different approaches and methodology. We randomly assigned twelve premature infants(gestational age < 32 week) at high risk of requiring erythrocyte transfusion for anemia of prematurity with either subcutaneous recombinant human erythropoietin or a placebo. Treatment with rHuEPO was initiated at a dose of 100 units/kg day for 3 days a week. All patients were given supplemental oral iron therapy at a dose of 3 mg/kg per day, as tolerated and oral vitamin E at a dose of 25 units per day. Treated and control babies did not differ with respect to weight, hematocrit, overall mean reticulocyte count or rate of growth respectively. However, reticulocyte counts increased earlier in patients given rHuEPO. We conclude that rHuEPO administration is safe and feasible at the dose studied.