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Monoclonal Antibody CFC-6, which Binds to Helix II, Inhibits Erythropoietin-Induced Bioactivity
Oh,Myung-Suk,Park,Ji-Sook,Ha,Byung-Jhip,Yoo,Ook-Joon,Yoo,Ree-Ann,Lee,Dong-Eok,Kim,Suk-Joon,Woo,Koo,Kim,Hyun-Su The Korea Science and Technology Center 1997 BMB Reports Vol.30 No.5
It was discovered that monoclonal anti-erythropoietin (EPO) antibody CFC-6 can neutralize EPO-induced cell activation. To know the binding site of CFC-6, recombinant human erythropoietin (rhEPO) was digested with Glu-C, followed by a separation using high performance liquid chromato graphy (HPLC). Each HPLC fraction was blotted on the nitrocellulose membrane and the membrane was treated with anti-EPO antibody CFC-6 and anti-mouse antibody which is modified with peroxidase. Only one spot showed the color and the fraction was sequenced by Edman degradation. The result suggest that CFC-6 recognizes amino acid sequence V63-W-Q-G-L-A-L-L-S-E72 which is a part of helix Ⅱ of the EPO molecule. Binding of CFC-6 to EPO may inhibit EPO binding to its receptor, which implies that the antibody binding site and the receptor binding site are close or overlapping.
만성신부전증 환자에서 Recombinant Human Erythropoietin 치료에 따른 혈액지표와 Hemoglobin A1c에 대한 연구
이세영,배성한,변동원,서교일,유명희,김극배 순천향의학연구소 1996 Journal of Soonchunhyang Medical Science Vol.2 No.2
Hemoglobin A1c is produced by a progressive, non-enzymatic reaction between glucose and hemoglobin within the erythrocytes. The HbA1c concentration is dependent on the plasma glucose level and the stage of development of the erythrocytes. Immature erythrocytes contain lower levels of glycosylated hemoglobin than mature erythrocytes. HbA1c level was decreased in short RBC life span. Therefore, HbA1c level is not only measure for assessment of moderate to long term glycemic status in diabetics, but also as a possible diagnostic parameter of anemia. The aim of this study was to evaluate the HbA1c level in patients with chronic renal failure with anemia. HbA1c concentration, iron, ferritin, TIBC and hematologic parameters were measured before treatment and 1, 2 months after administration of recombinant human erythropoietin. The HbA1c concentration was measured by high performance liquid chromatography with cation exchange column (Pharmacia). The results were as follows; 1. The mean of hemoglobin was 8.66 g/dL in the controls and 7.84 g/dL in the patients with chronic renal failure. The mean of hematocrit was 25.19 % in the controls and 23.14 % in the patients with chronic renal failure. The mean of MCV was 93.23 fL in the controls and 92.73 fL in the patients with chronic renal failure. The mean of MCH was 32.56 pg in the controls and 31.76 pg in the patients with chronic renal failure. The mean of HbA1c was 3.15 % in the controls and 2.95 % in the patients with chronic renal failure. 2. Hematologic parameters in the patients with chronic renal failure 1) The results of hemoglobins were 7.84 g/dL, 8.11 g/dL, 8.92 g/dL, the MCH were 31.76 pg, 32.87 pg, 33.20 pg, the results of MCV were 92.73 fL, 97.37 fL, 92.85 fL at before treatment and 1, 2 months after administration of r-HuEpo. 2) The results of hematocrits were 23.14 %, 23.73 %, 26.73 % at before treatment and 1, 2 months after administration of r-HuEpo. 3. Iron metabolism parameters in the patients with chronic renal failure 1) The results of iron test were 180.92 ㎍/dL, 137.79 ㎍/dL, 126.83 ㎍/dL at before treatment and 1, 2 months after administration of r-HuEpo. 2) The results of ferritin test were 1500.2 ng/ml, 1311.6 ng/ml, 1151.0 ng/ml at before treatment and 1, 2 months after administration of r-HuEpo. 3) The results of TIBC test were 282.17 ㎍/dL, 282.45 ㎍/dL, 278.83 ㎍/dL at before treatment and 1, 2 months after administration of r-HuEpo. 4. The results of HbA1c test were 2.95 %, 3.08 %, 3.18 % at before treatment and 1, 2 months after administration of r-HuEpo. It is suggested that, in patients with chronic renal failure, evaluation of HbA1c in diabetics who have anemia with chronic renal failure should be consider possible hematologic parameters, and HbA1c level would be one of the marker of anemia status, but further studies are needed.
Monoclonal Antibody CFC-6, which Binds to Helix II, Inhibits Erythropoietin-Induced Bioactivity
Ha, Byung-Jhip,Kim, Suk-Joon,Park, Ji-Sook,Yoo, Ree-Ann,Lee, Dong-Eok,Yoo, Ook-Joon,Woo, Koo,Kim, Hyun-Su,Oh, Myung-Suk Korean Society for Biochemistry and Molecular Biol 1997 Journal of biochemistry and molecular biology Vol.30 No.5
It was discovered that monoclonal anti-erythropoietin (EPO) antibody CFC-6 can neutralize EPO-induced cell activation. To know the binding site of CFC-6, recombinant human erythropoietin (rhEPO) was digested with Glu-C, followed by a separation using high performance liquid chromato graphy (HPLC). Each HPLC fraction was blotted on the nitrocellulose membrane and the membrane was treated with anti-EPO antibody CFC-6 and anti-mouse antibody which is modified with peroxidase. Only one spot showed the color and the fraction was sequenced by Edman degradation. The results suggest that CFC-6 recognizes amino acid sequence V63-W-Q-G-L-A-L-L-S-E72 which is a part of helix II of the EPO molecule. Binding of CFC-6 to EPO may inhibit EPO binding to its receptor, which implies that the antibody binding site and the receptor binding site are close or overlapping.
빈혈을 동반한 만성신부전에서 Recombinant Human Erythropoietin 의 효과
이경생(Kyoung Saeng Lee),이인생(In Saeng Lee),한수용(Su Yong Han),추장식(Jang Sik Choo),서상렬(Sang Yeol Suh),송창섭(Chang Sup Song) 대한내과학회 1993 대한내과학회지 Vol.45 No.1
Background: Anemia is a major complication of chronic renal failure. Recombinant human erythropoietin (rhuEPO) has proven to be a highly effective treatment for the anemia of end-stage renal disease. Methods: We administered rhuEPO to 15 anemic patients of end-stage renal disease who were undergoing hemodialysis. Recombinant human erythropoietin was given intravenously twice a week after dialysis. Transfusion requirements, hemoglobin, hematocrit, reticulocyte, ferrokinetics, BUN, creatinine, electrolytes and adverse reactions were monitored. Results: 1) In 15 patients treated for 8 weeks, hemoglobin level increased from initial mean (±SD) 6.4±0.64 g/dl to 8.6±0.69g/df (p<0.01). Hematocrit level increased from initial mean 18.9±1.79% to 25.2±3.36% (p<0.01). Reticulocyte level increased from initial mean 1.2±0.93% to 2.1±1.02% (p<0.01). 2) Serum iron level decreased from initial mean 128.3±98.54 ㎍/dl to 95.4±75.41 ㎍/dl (p<0.01) and ferritin level decreased from initial 1454.3±732.52±52 ng/dl to 1213.5±534.43 ng/dl (p<0.05). 3) There were no significant differences in the serum levels of BUN, creatinine, Na K and phosphorus before and after the treatment. 4) Adverse reactions of rhuEPO were observed in 2 patients. One was occlusion of arteriovenous fistula and the other was severe itching sensation. 5) The patients were transfused 1.6 pints of packed red cell per month before the treatment, but they no longer needed transfusion after the treatment. Conclusion: These results suggest that recombinant human erythropoietin is safe and may eliminate the need for transfusion in anemic patients of end-stage renal disease.
투석치료를 받는 만성신부전 환자의 빈혈에 대한 유전자 재조합 인 에리트로포이에틴(에스포젠)의 임상효과
안규리(Curie Ahn),오하영(Ha Young Oh),박정식(Jung Sik Park),정우경(Woo Kyung Chung),호지숙(Gee Suk Ho),허우성(Woo Sung Huh) 대한신장학회 2000 Kidney Research and Clinical Practice Vol.19 No.2
N/A We conducted a multicenter clinical trial to evaluate the efficacy and safety of recombinant human erythropoietin(Espogen, LG Chemical Ltd.) in the anemic patients of chronic renal failure undergoing dialysis. The patients were end-stage renal disease who were undergoing hemodialysis or peritonea1 dialysis for 3 months or longer and they had less than 8g/dL of hemoglobin and more than 100ng/mL of serum ferritin. Hemodialysis patients were administered 150unit/kg/week of recombinant human erythropoietin as initial dose, and peritoneal dialysis patients 50unitAg, twice per week. We examined hemoglobin value every other week and adjusted the dose in order to maintain hemoglobin level as 10-llg/dL. We enrolled 64 patients and analysed 54 cases in the final. 96.3%(52/54) of patients showed increase by more than 1.0g/dL and the others in- crease by more than 0.5g/dL. Baseline hemoglobin, hematocrit were 7.11±0.85g/dL, 21.3±2.6% and final level were 10.42±1.31g/dL, 31.9±3.5%(p=0.0001), respectively. Reticulocyte was increased after 2 weeks of administration from 0.90±0.74% to 2.45±0.84% The adverse effects included hypertension, headache, increased potassium and phosphate level so required regular monitoring. Therefore we showed that Es-pogen was effective in correcting the anemia of chronic renal failure and didn't have any particular adverse effects.
DA-3585(recombinant human erythropoietin)의 국소자극성에 관한 연구
조현,김동환,강경구,박장현,이성희,김원배,Cho, Hyeon,Kim, Dong-Hwan,Kang, Kyung-Koo,Park, Jang-Hyeon,Lee, Sung-Hee,Kim, Won-Bae 한국독성학회 1998 Toxicological Research Vol.14 No.3
As a series of safety studies on DA-3585, a recombinant human erythropoietin, its local irritancy was examined in rabbits after the following treatments; application into the conjunctival sac of the eye(single), subcutaneous injection (single and -day repeated)and intravenous injection (7-day repeated.)In addition, perivascular irritation of DA-3585 was investigated in mice. In the result of ocular irritation test, 10,000IU/ml solution of DA-3585 could be considered as a non-irritating material. The local irritation of DA-3585 by a single and 7-day repeated subcutaneous injection was negligible and not so much different from that of saline. In the vascular irritancy test, macro-and microscopic observations revealed that local irritation of DA-3585 was comparable to that of saline when injected into retroauricular vein of rabbits for 7 consecutive days. Furthermore the perivascular administration of DA-3585 upto the concentration of 10,000 IU/ml did not induce any morphological abnormalities at injection sites. The results obtained from the present study suggest that the local irritancy of DA-3585 is not different from that of saline when injected through intravenous or subcutaneous route for clinical practice.
비글개에서 인체 재조합 적혈구 조혈인자, rHu-EPO의 급성독성에 관한 연구
조명행,성하정,김형식,곽승준,천선아,임소영,김원배,김병문,안병옥,이병무,Cho, Myung-Hang,Seong, Ha-Jung,Kim, Hyung-Sik,Kwack, Seung-Jun,Chun, Sun-Ah,Lim, So-Young,Kim, Won-Bae,Kim, Byoung-Moon,Ahn, Byoung-Ok,Lee, Byung-Mu 한국독성학회 1996 Toxicological Research Vol.12 No.2
The acute toxicity of rHu-EPO, newly developed recombinant erythropoietin, was tested in beagle dogs. rHu-EPO, when administered intravenously at 25, 000 IU/kg, did not cause any death. Also, rHu-EPO did not induce any change of body weight, food intake and clinical signs compared to controls. There were no significant changes in hematological, urine analysis and pathological examination. These results showed that rHu-EPO did not induce any remarkable toxic response and the $LD_50$ was greater than 25, 000 IU/kg in beagle dogs.