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 $0.1-1.1{\times}10^9M^{-1}$.

포병 표적탐지 레이더 운용의 계량적 효과 분석

강신성,이재영,Kang, Shin-Sung,Lee, Jae-Yeong 한국시뮬레이션학회 2010 한국시뮬레이션학회 논문지 Vol.19 No.2

In the future warfare, the importance of the counter-fire operation is increasing. The counter-fire operation is divided into offensive counter-fire operation and defensive counter-fire operation. Reviewing the researches done so far, the detection asset of offensive counter-fire operation called UAV(Unmanned Aerial Vehicle) and its operational effectiveness analysis is continually progressing. However, the analysis of the detection asset of defensive counterfire called Target Acquisition Radar(TAR) and its quantitative operational effectiveness are not studied yet. Therefore, in this paper, we studied operational effectiveness of TAR using C2 Theory & MANA Simulation model, and showed clear quantitative analysis results by comparing both cases of using TAR and not using TAR.

시뮬레이션을 이용한 표적탐지 레이더 운용효과에 관한 연구

강신성(Shin-sung Kang),이재영(Jae-Yeong Lee) 한국경영과학회 2009 한국경영과학회 학술대회논문집 Vol.2009 No.10

In the future warfare, the importance of the counter fire operation is increasing. The counter-fire operation is divided into offensive counter-fire operation and defensive counter-fire operation. Reviewing the researches done so far, the detection asset of offensive counter-fire operation called UAV(Unmanned Aerial Vehicle) and its operational effectiveness is continually progressing. However, the analysis of the detection asset of defensive counter-fire called Target Acquisition Radar and its quantitatively operational effectiveness are currently none. The research we have seen so far were done through the operational effectiveness of Target Acquisition Radar, brought into the ROK-Army, MANA by modeling quantitatively and analyzing them.

펩티드성분을 함유하는 블록공중합체막의 세포접착성

강인규,강신성,임학상,Kang, Inn-Kyu,Kang, Shin-Sung,Lim, Hak-Sang 대한의용생체공학회 1992 의공학회지 Vol.13 No.2

Attachment and growth of mouse flbroblast cells on block copolypeptides were studied in the pres once or absence of serum proteins. Cells are rapidly attached to she polymer surface within 30 min regardless of substrate in the presence of serum. The number of flbroblast cells attached on the poly mer surface coated by collagen was larger than thats on the bare surface. Attachment of cells Is as a whole suppressed to a low level by the addltion of sodium azide in the absence of serum. Thls suE gests that the active attachment of cells requires the biological metabolism taking place on polymer substrates. In the presence of serum protein, flbroblast cells are more rapidly grown on the bolck co polymer consisting of poly(T-benzyl L-glutamate) (PBLG) and polyoxypropylene(POP) than on other block copolymers. These results were in agreement wish the data obatlned by an Inverted ml croscope.

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.

Human Immunodeficiency Virus Type 1 의 Rev Responsive Element 의 기능 분석

이형열,이안휘,강신성,성영철 ( Hyeong Yeol Lee,Ann Hwee Lee,Shin Sung Kang,Young Chul Sung ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.3

Expression of the structural proteins of human immunodeficiency virus type (HIV-1) requires the Rev protein encoded by the rev open reading frame. Rev protein interacts with Rev-responsive element RRE located in the env region of the viral mRNA and seems to mediate the export of the incompletely spliced viral mRNA to the cytoplasm. RRE has a complex secondary structure which is composed of a central stem (I`), a small stem (I) and 5 stem/loops (II, III, IV, V, VI). To investigate which region of RRE is essential for the interaction with Rev protein, mutational analysis in RRE was carried out. We examined the nature of the mutated RRE in several assay systems, p24 ELISA assay, Reverse Transcriptase assay and CAT assay. Here we show that the secondary structure of stem/loop II region is critical for Rev response. Other structural components within RRE RNA seem to have only subsidiary roles. We also show that RRE appears to contain a negative sequence which hinders the expression of structural gene in the absence of the Rev protein.

Human Immunodeficiency Virus Type 1의 Rev-Responsive Element의 기능 분석

이형열,이안휘,강신성,성영철,Lee, Hyeong-Yeol,Lee, Ann-Hwee,Kang, Shin-Sung,Sung, Young-Chul 생화학분자생물학회 1992 한국생화학회지 Vol.25 No.3

Human immunodeficiency virus type 1(HIV-1)의 Rev 단백질은 자신의 복제에 필수적으로 작용하는 조절단백질이다. 이 단백질은 구조유전자 mRNA를 핵에서 세포질로의 수송을 촉진함으로서 구조단백질의 발현을 유도하는 것으로 생각된다. 이러한 Rev 단백질은 HIV-1 RNA의 env 유전자 내에 존재하는 cis-acting element인 Rev responsive element(RRE)와 직접 결합함으로써 그 기능을 나타낸다. 컴퓨터 분석결과 RRE는 복잡한 RNA 이차구조 즉 하나의 central stem (I')과 작은 stem(I), 그리고 5개의 stem/loop들로(II, III, IV, V, VI) 구성되는 것으로 예견 되어졌다. 본 연구에서는 RRE RNA내의 어느 영역이 Rev 조절단백질의 반응에 필수적으로 작용하는가를 알아보기 위하여 RRE RNA내의 안정한 이차구조를 in vitro mutagenesis시켜 Rev 단백질과 RRE RNA mutants와의 상호작용을 p24 ELISA, reverse transcriptase assay, CAT assay 등의 방법으로 살펴보았다. 그 결과 RRE의 복잡한 이차구조 내에서 stem/loop II의 이차구조가 Rev 단백질의 작용에 필수적임을 밝혔으며, 다른 영역의 이차구조들은 Rev 조절단백질과 stem/loop II와의 상호작용을 증가시킴을 관찰하였다. 또한, stem/loopII내의 stem에는 Rev 단백질과의 반응에 관련된 기능 외에 Rev 단백질이 없을 때 구조유전자의 발현을 억제하는 기능을 가진 부위, 즉 cis-acting repressive sequence(CRS)가 존재함을 확인하였다. Expression of the structural proteins of human immunodeficiency virus type (HIV-1) requires the Rev protein encoded by the rev open reading frame. Rev protein interacts with Rev-responsive element RRE located in the env region of the viral mRNA and seems to mediate the export of the incompletely spliced viral mRNA to the cytoplasm. RRE has a complex secondary structure which is composed of a central stem (I'), a small stem (I) and 5 stem/loops (II, III, IV, V, VI). To investigate which region of RRE is essential for the interaction with Rev protein, mutational analysis in RRE was carried out. We examined the nature of the mutated RRE in several assay systems, p24 ELISA assay, Reverse Transcriptase assay and CAT assay. Here we show that the secondary structure of stem/loop II region is critical for Rev response. Other structural components within RRE RNA seem to have only subsidiary roles. We also show that RRE appears to contain a negative sequence which hinders the expression of structural gene in the absence of the Rev protein.

Immune Response of BALB/c and Production of Monoclonal Antibodies against Outer Membrane Proteins from Vibrio vulnificus

Han-Do Kim(김한도),Shin-Sung Kang(강신성),Jin-Woo Ju(주진우) 大韓微生物學會 1988 大韓微生物學會誌 Vol.23 No.3