사람 α - Proteinase Inhibitor 의 분리 및 분석

방옥선,최윤정,강신성 ( Ok Sun Bang,Yoon Jeong Choi,Shin Sung Kang ) 생화학분자생물학회 1988 BMB Reports Vol.21 No.4

For the preliminary step to make and characterize the monoclonal antibodies against human alpha-1-proteinase inhibitor (α₁-PI), the α₁-PI molecules were isolated from 500 ml of normal human serum through the procedures including salt precipitation, Cibacron blue F3GA-agarose, DEAE-cellulose, concanavalin A-Sepharose and Sephadex G-100 gel chromatography. The isolated α₁-PI proparation showed a single band both on polyacrylamide gel electrophoresis and SDS-polyacrylamide gel electrophoresis and also a single precipitin arc against anti-normal human serum and anti-human α₁-PI antiserum on immunoelectrophoresis. The recovery of α₁-PI was 52.42 ㎎ total and the specific activity of this protein was 14.29 unit/㎎ of protein. The CNB-cleaved peptide fragments (Fr I : 64-220; Fr II : 243-351; Fr III : 1-63) were also isolated to homogeneity.

사람 ${\alpha}_1$-Proteinase Inhibitor 의 분리 및 분석

방옥선,최윤정,강신성,Bang, Ok-Sun,Choi, Yoon-Jeong,Kang, Shin-Sung 생화학분자생물학회 1988 한국생화학회지 Vol.21 No.4

사람의 ${\alpha}_1$-proteinase inhibitor (${\alpha}_1-PI$)에 대한 단일클론 항체의 생산 및 분석을 위하여 사람의 혈장을 재료로 추출법, Cibacron blue F3GA-agarose, DEAE-cellulose, Concanavalin A-Sepharose 4B, Sephadex G-100 등의 크로마토그라피 방법으로 ${\alpha}_1-PI$를 분리하였다. 분리 한 ${\alpha}_1-PI$는 alkaline polyacrylamide gel electrophoresis 및 SDS-PAGE 에서 단일 밴드로 나타났고, 또 immunoelectrophoresis에서도 anti-human serum 및 anti-${\alpha}_1-PI$에 대해 단일 침전 arc를 나타내어 순수분리되었음을 확인하였다. 정상인 혈장 500 ml로부터 분리된 ${\alpha}_1-PI$의 회수율은 약 37 %로서 52.41 mg이었고, 트립신 저해능의 비활성도는 14.29 unit/mg protein이었다. 이와같이 하여 순수정제된 ${\alpha}_1-PI$를 CNBr로 처리하여 3 가지의 $CNBr^-$ 펩티드 분획인 FrI (아미노산 잔기 64-220), Fr II (아미노산 잔기 243-351) 및 Fr III (아미노산 잔기 1-63)을 각각 분리하였다. For the preliminary step to make and characterize the monoclonal antibodies against human alpha-I-proteinase inhibitor (${\alpha}_1-PI$), the ${\alpha}_1-PI$molecules were isolated from 500 ml of normal human serum through the procedures including salt precipitation, Cibacron blue F3GA-agarose, DEAE-cellulose, concanavalin A-Sepharose and Sephadex G-100 gel chromatography. The isolated ${\alpha}_1-PI$ proparation showed a single band both on polyacrylamide gel electrophoresis and SDS-polyacrylamide gel electrophoresis and also a single precipitin arc against anti-normal human serum and anti-human ${\alpha}_1-PI$) antiserum on immunoelectrophoresis. The recovery of $a_1$-PI was 52.42 mg total and the specific activity of this protein was 14.29 unit/mg of protein. The CNB-c1eaved peptide fragments (Fr I: 64-220: Fr II: 243-351; Fr III: 1-63) were also isolated to homogeneity.

단핵구 분화에 있어서 fibronectin 수용체의 역할

방옥선(Ok Sun Bang),강희갑(Hee Kap Kang),박의균(Eui Kyun Park),이영섭(Young Sup Lee),강신성(Shin Sung Kang) 경북대학교 생명공학연구소 1994 遺傳工學硏究所報 Vol.9 No.1

N/A

단핵구 분화에 대한 Fibronectin 및 그 단편의 역할

방옥선(Ok Sun Bang),이영섭(Young Sup Lee),강신성(Shin Sung Kang) 경북대학교 생명공학연구소 1993 遺傳工學硏究所報 Vol.8 No.1

N/A

하이브리도마 방법에 의한 항 α - Proteinase Inhibitor 모노클론항체의 생산과 분석

강신성,방옥선,강희갑,손종경 ( Shin Sung Kang,Ok Sun Bang,Hee Kap Kang,Jong Kyung Sonn ) 생화학분자생물학회 1988 BMB Reports Vol.21 No.4

Monoclonal antibodies (MAbs) against human α₁-PI was produced by hybridizing SP 2/0 -Ag 14 mouse myeloma cells with spleen cells of Balb/c mice immunized with a α₁-PI. The resulting hybridoma clones were screened for their ability to bind α₁-PI by ELISA. Seven positive stable monoclones were obtained by cloning three times in serial dilutions, and each antibody was identified to be homogeneous by several criterial. By western bloting analysis using CNBr-cleavage peptide fragments (Fr I: amino acid residues 64-220; Fr II: 243-351; Fr III : 1-63), four of these monoclones were found to synthesize MAb specific to Fr I, and one clone to Fr II. But MAbs of the other two clones showed reactivity only to intact α₁-PI. Five of our monoclones were appeared to synthesize IgG₁(k) antibodies having affinity constant in the range 0.1-1.1×10^9 M^(-1)

Production and Characterization of Monoclonal Antibodies against Human alpha-1-Proteinase Inhibitor by Hybridoma

강신성,방옥선,강희갑,손종경,Kang, Shin-Sung,Bang, Ok-Sun,Kang, Hee-Kap,Sonn, Jong-Kyung 생화학분자생물학회 1988 한국생화학회지 Vol.21 No.4

Alpha-1-proteinase inhibitor(${\alpha}_1-PI$)로 면역시킨 Balb/c 마우스 비장세포와 SP 2/0-Ag 14 마우스 미에로마 세포를 융합시키는 하이브리도마 기법으로 모노클론 항체 생산을 실시하였다. 하이브리드클론들이 생산하는 항체들과 ${\alpha}_1-PI$와의 반응성은 ELISA로 분석하였으며, 양성클론을 제한희석법으로 클로닝하여 7클론의 항-${\alpha}_1-PI$ 특이 클론세포를 얻었고, 이들의 단일클론성을 확인하였다. 7클론 중 5클론이 생산하는 모노클론항체는 항원에 대한 결합상수가 $0.1-1.1{\times}10^9M^{-1}$의 $IgG_1(k$) 이었고, 이들 항체의 항원 특이성을 ${\alpha}_1-PI$의 CNBr-peptide 분획을 이용한 Western blot를 실시한 결과 FrI(아미노산 잔기 64-220)에 특이한 것이 4클론, Fr II (243-351)에 특이한 것이 1클론이었고, 나머지 2클론의 특이성은 결정할 수 없었다. Monoclonal antibodies (MAbs) against human ${\alpha}_1-PI$ was produced by hybridizing SP 2/0-Ag 14 mouse myeloma cells with spleen cells of Balb/c mice immunized with ${\alpha}_1-PI$. The resulting hybridoma clones were screened for their ability to bind ${\alpha}_1-PI$ by ELISA. Seven positive stable monoclones were obtained by cloning three times in serial dilutions, and each antibody was identified to be homogeneous by several criterial By western bloting analysis using CNBr-cleavage peptide fragments (Fr I: amino acid residues 64-220; Fr II: 243-351; Fr III: 1-63), four of these monoclones were found to synthesize MAb specific to Fr I, and one clone to Fr II. But MAbs of the other two clones showed reactivity only to intact $a_1$-PI. Five of our monoclones were appeared to synthesize $IgG_1(k$) antibodies having affinity constant in the range $0.1-1.1{\times}10^9M^{-1}$.

Human Paraoxonase 1(PON1)의 유전자 다형성에 따른 중풍환자의 습담 변증과의 상관성 연구

임지혜,고미미,이정섭,방옥선,차민호,Lim, Ji-Hye,Ko, Mi-Mi,Lee, Jung-Sup,Bang, Ok-Sun,Cha, Min-Ho 대한한방내과학회 2010 大韓韓方內科學會誌 Vol.31 No.4

Objective : In the present study, we investigated genetic distribution of eight single nucleotide polymorphisms of PON1 between Dampness and Phlegm and non Dampness and Phlegm pattern identification(PI) among Korean stroke patients. Materials and Methods : One hundred forty stroke subject without Dampness and Phlegm and fifty eight stroke subjects with Dampness and Phlegm were participated in this study. After informed consents, eight single nucleotide polymorphisms(SNPs) in PON1 of each subjects were identified by DNA sequencing and primer extension method and statistical analysis was performed to determine the significant difference between Dampness and Phlegm and non Dampness and Phlegm groups. Results : Among anthropometric characteristics and blood parameters, waist circumference and total cholesterol were significantly higher in Dampness and Phlegm. Among 8 SNPs of PON1, frequency of M allele and subjects with M allele in L55M SNP were significantly higher in Dampness and Phlegm group (p=0.0032 and p=0.0053, respectively) but subjects with T allele in C-2033T SNP were lower in Dampness and Phlegm group(p=0.0302). Effect of L55M and C-2033T on Dampness and Phlegm were 3.07% and 1.75%, respectively. Conclusion : Our results suggest that L55M SNP in exon and C-2033T in promoter region of PON1 maybe affect to Dampness and Phlegm pattern identification. However, further study should be carried out to find out the detailed mechanism how L55M and C-2033T can affect Dampness and Phlegm stroke patients.

뇌졸중 위험지표로서의 일반혈액검사 소견에 대한 환자;대조군 연구

이현의,강경원,유병찬,방옥선,백경민,설인찬,김윤식,Lee, Hyon-Ui,Kang, Kyung-Won,Yu, Byeong-Chan,Bang, Ok-Sun,Baek, Kyung-Min,Seol, In-Chan,Kim, Yoon-Sik 대한한방내과학회 2007 大韓韓方內科學會誌 Vol.28 No.4

Objective : Stroke is one of the most common causes of death in Korea. This study was done to evaluate the association of complete blood count (CBC) with the risk of hemorrhagic stroke and ischemic stroke. Methods : In 217-case patients with ischemic stroke or hemorrhagic stroke and 146 healthy control subjects without stroke, hypertension, diabetes mellitus, hyperlipidemia, or ischemic heart disease and 160 controls without ischemic stroke or hemorrhagic stroke, we tested and compared white blood cell count (WBC), red blood cell count (RBC), hemoglobin (Hgb), hematocrit (Hct) and platelet. These data were statically analyzed by general linear models and binary logistic regression analysis to get each adjusted odds ratio. Results :The level of WBC was significantly higher in all cases. The level of RBC, Hct and Hgb was significantly lower in patients of ischemic stroke. The level of platelet was significantly higher in patients of ischemic stroke. Conclusion : These results suggest high WBC may be a risk factor of hemorrhagic stroke and ischemic stroke and low RBC, low Hct, low Hgb and high platelet may be risk factors of ischemic stroke in Koreans.

무증상 뇌경색의 위험요인에 대한 환자;대조군 연구

백혜기,고미미,유병찬,방옥선,오영선,김연진,김정현,김윤식,설인찬,Baek, Hye-Ki,Ko, Mi-Mi,Yu, Byeong-Chan,Bang, Ok-Sun,Oh, Yeong-Seon,Kim, Yeon-Jin,Kim, Jung-Hyun,Kim, Yoon-Sik,Seol, In-Chan 대한한방내과학회 2007 大韓韓方內科學會誌 Vol.28 No.4

Background : Cerebrovascular disease is a major cause of death and disability in adults. Silent cerebral infarction (SCI) portends more severe cerebral infarction or may lead to insidious progressive brain damage resulting in vascular dementia. Known cardiovascular risk factors, such as arterial hypertension, diabetes mellitus, smoking, hyperlipidemia and ischemic heart disease may increase the risk of SCI. This study was designed to evaluate the risk factors of SCI in an apparently normal adult population. Methods : We divided 340 neurologically normal adults (mean age=59.90$\pm$8.30, men:women = 146:194) who underwent brain computed tomography (CT) or magnetic resonance imaging (MRI) at the Stroke Medical Center in Daejeon University Oriental Medicine Hospital in two groups, Silent inf. and Controls,and analyzed risk factors of SCI by interview, physical examination and blood test. Risk factors of SCI were assessed by interview, physical examination and blood test. We performed Pearson's chi-square test and two-sample t-test for univariate analysis and multiple logistic regressions for multivariate analysis to evaluate risk factors of SCI. Results : Old age, diabetes mellitus, and high lactate dehydrogenase (LDH) levels were associated with SCI on univariate analysis. Diabetes mellitus was demonstrated to be an independent risk factor for SCI on multivariate analysis. Conclusions : Advanced age, diabetes mellitus, and LDH levels are associated with SCI.

급성기 중풍환자에서 비만 및 혈액지표의 기허 및 화열 변증의 차이에 대한 고찰

차민호,김소연,임지혜,강병갑,고미미,김노수,이정섭,방옥선,Cha, Min-Ho,Kim, So-Yeon,Lim, Ji-Hye,Kang, Byung-Kab,Koo, Mi-Mi,Kim, No-Soo,Lee, Jeong-Sub,Bang, Ok-Sun 대한한방내과학회 2009 大韓韓方內科學會誌 Vol.30 No.4

Object : In the present study, we investigated the obesity and blood parameters between Qi-deficiency and Fire/Heat pattern identification/syndrome differentiation (PI/SD) in acute stroke patients. Materials and Methods : A total of 391 stroke patients within 7 days after onset were consecutively recruited from 12 hospitals across South Korea from Nov. 1st, 2006 to Jun. 31st, 2009. They were diagnosed as Fire/Heat or Qi-deficiency among five PI/SD subtypes by two independent stroke experts. We investigated the differences of obesity and blood characteristics between Fire/Heat and Qi-deficiency by statistical analyses. Results : In male subjects, obesity was significantly associated with Fire/Heat PI/SD. The averaged mean BMI ($24.13kg/m^2$) and waist circumference(89.34cm) of the Fire/Heat group were higher than those of the Qi-deficiency group ($22.60kg/m^2$ and 83.43 cm, respectively). The number of obese patients was larger in the Fire/Heat group than in the Qi-deficiency group (p = 0.001). Hyperlipidemia was also related with Fire/Heat. However, obesity was not associated with PI/SD in female subjects where the number of hyperlipidemic patients was higher in the Qi-deficiency group. Among blood parameters, the levels of triglycerides and fasting blood sugar were higher in the Fire/Heat group compared with the Qi-deficiency group in male subjects. However, total cholesterol of the Qi-deficiency group was higher than in the Fire/Heat group among female subjects. Conclusion : This study shows that obesity and hyperlipidemia are significantly difference between Qi-deficiency and Fire/Heat. We suggests that PI/SD may be associated with clinical characteristics and large population study between PI/SD and clinical characteristics including blood parameters are needed.