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박희섭,정미영,오명석,고중환,김현수,현형환,Park, H.S.,Jung, M.Y.,Oh, M.S.,Koh, J.H.,Kim, H.S.,Hyun, H.H. 생화학분자생물학회 1987 한국생화학회지 Vol.20 No.4
유전자 조작된 대장균이 생산하는 알파 인터페론을 단일 항체 칼럼을 사용하여 효과적으로 정제하였다. rIFN-$\alpha$에 대한 항체를 생산하는 단일클론 융합세 포를 쥐(BALB/c)의 복강에 접종하여 얻은 복강액으로부터 Protein A column을 사용하여 항체를 정제하였다. 이 정제된 항체를 Affigel-10에 결합시켜 단일항체 칼럼을 만든 후, 유전자 조각 장균을 6M 구아니딘 (guanidine-HCl)으로 처리 하여 얻은 인터페론 포함 상등액을 통과시켜 인터페론을 정제하였다. 정제된 인터페론 $2{\times}10^8\;IU/mg$ protein의 specific activity를 가졌으며 분자량은 SDS-PAGE에서 18,600이었고 30개까지 이이노산 서열을 분석한 결과 천연의 human leukocyte IFN-$\alpha$의 것과 동일하였다. 비환원 조건에서 SDS-PAGE 한 결과 약 15%의 인터페론 이중체 (Interferon dimer)를 포함 하고 있었다. 이를 제거하기 위해 생리식염수 용액으로 사용하여 HPLC 젤여과를 행하였다. 최종 정제된 인터페론을 비환원 조건에서 SDS-PAGE 한 결과 인터페론 단일체의 순도는 99.9% 이상인 것으로 판명되었으며, 최종 정제된 인터페론은 약 5개의 등전 전위점을 갖고 있다 (pI = 6.12, 6.06, 5.85, 5.58, 5.30). Immunoadsorbent chromatography technique was applied for the efficient purification of Interferon (rIFN-$\alpha$) from recombinant E. coli. Monoclonal antibody to rIFN-$\alpha$ was purified using protein A-Sepharose CL-4B chromatography from ascites fluid of BALB/c mice which were innoculated with monoclonal hybridoma cell. The purified antibody was coupled to Affigel-10, and used as an immunoadsorbent for IFN. The recombinant E. coli cells were sedimented and cell-opening was done using 6 M guanidine-HCl. It was loaded to the above immunoadsorhent column and then the bound IFN was eluted from the column with 0.2 M acetic acid. As a result of purification, the specific activity of the purified IFN was $2{\times}10^8\;IU/mg$ protein and the purification fold was 4100. The amino-terminal sequence of purified IFN was exactly identical to that of natural human leukocyte IFN upto 30 amino acid residues except the 1st and 29th unidentified amino acid residues. The molecular weight of the purified IFN was estimated to be 18,600 by SDS-PAGE. Since IFN purified by monoclonal antibody column contains IFN dimer up to 15%, further purification was done by HPLC gel filtration. The purity of finally-purified IFN monomer was estimated to be more than 99.9% by SDS-PAGE under non-reducing condition. Finally-purified IFN monomer has several pI points (6.12, 6.06, 5.85, 5.58, and 5.30).
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단일항체 칼럼과 고압 액체크로마토그래피에 의한 유전자 조작 사람 알파 인터페론의 분리 및 정제
박희섭,정미영,오명석,고중환,김현수,현형환 ( H . S . Park,M . Y . Jung,M . S . Oh,J . H . Koh,H . S . Kim,H . H . Hyun ) 생화학분자생물학회 1987 BMB Reports Vol.20 No.4
Immunoadsorbent chromatography technique was applied for the efficient purification of Interferon (rIFN-α) from recombinant E. coli. Monoclonal antibody to rIFN-α was purified using protein A-Sepharose CL-4B chromatography from ascites fluid of BALB/c mice which were innoculated with monoclonal hybridoma cell. The purified antibody was coupled to Affigel-10, and used as an immunoadsorbent for IFN. The recombinant E. coli cells were sedimented and cell-opening was done using 6 M guanidine-HCl. It was loaded to the above immunoadsorbent column and then the bound IFN was eluted from the column with 0.2 M acetic acid. As a result of purification, the specific activity of the purified IFN was 2 × 10^8 IU/㎎ protein and the purification fold was 4100. The amino-terminal sequence of purified IFN was exactly identical to that of natural human leukocyte IFN upto 30 amino acid residues except the lst and 29th unidentified amino acid residues. The molecular weight of the purified IFN was estimated to be 18,600 by SDS-PAGE. Since IFN purified by monoclonal antibody column contains IFN dimer up to 15% further purification was done by HPLC gel filtration. The purity of finally-purified IFN monomer was estimated to be more than 99.9% by SDS-PAGE under non-reducing condition. Finally-purified IFN monomer has several pI points (6.12, 6.06, 5.85, 5.58, and 5.30).
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이주원(J. W. Lee),최성수(S. S. Choi),김종우(J. W. Kim),정미영(M. Y. Jung),김대욱(D. W. Kim) 한국진공학회(ASCT) 2000 Applied Science and Convergence Technology Vol.9 No.2
Submicron aperture 제작 기술은 near field optical sensor 또는 liquid metal ion source에 응용될 수 있는 가능성으로 인해 흥미를 모으고 있다. 본 실험에서는 submicron aperture 제작에 대해 기술할 것이다. 먼저, 2 ㎛ 크기의 dot array를 광학 리소그라피 방법으로 패턴화하였다. KOH 비등방성 식각 방법으로 V-groove 형을 만든 후, 1000℃에서 600분 동안 건식 산화 작업을 거쳤다. 이 산화 과정에서 결정 방향에 따라 산화율이 달라지게 되는데 Si(111) 면은 Si(100)면에 비해 산화율이 커서 두꺼운 산화막이 형성되며, 이 막은 연이은 건식식각 과정에서 etch-mask로 활용된다. Reactive ion etching은 ICP (Inductively Coupled Plasma) 장비를 사용하였으며, V-groove의 바닥에 형성된 90 ㎚ 두께의 SiO₂와 그 아래의 Si을 식각하였다. 이 때, 기판에 걸린 negative bias는 Cl₂ RIE의 anisotropic etching 효과를 증대시키는 것 같았으며, SEM 촬영 결과 식각 후에 Si(111)면 위에는 약 130 ㎚ 정도의 산화층이 잔류하고 있었다. 이렇게 형성된 Si aperture는 향후 NSOM sensor 등에 적용될 수 있을 것이다. The fabrication of the submicron size hole has been interesting due to the potential application of the near field optical sensor or liquid metal ion source. The 2 micron size dot array was photolithographically patterned. After formation of the V-groove shape by anisotropic KOH etching, dry oxidation at 1000℃ for 600 minutes was followed. In this procedure, the orientation dependent oxide growth was performed to have an etch-mask for dry etching. The reactive ion etching by the inductively coupled plasma (ICP) system was performed in order to etch ~90 ㎚ SiO₂ layer at the bottom of the V-groove and to etch the Si at the bottom. The negative ion energy would enhance the anisotropic etching by the Cl₂ gas. After etching, the remaining thickness of the oxide on the Si(111) surface was measured to be ~130 ㎚ by scanning electron microscopy. The etched Si aperture can be used for NSOM sensor.
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김지선,노자민,류진영,오정연,이서주,정미영,조재실,주한별 이화여자대학교 간호과학대학 2011 이화간호학회지 Vol.- No.45
The purpose of this study was to investigate the aspect and degree of female university students' menstruation discomfort and to study how the menstruation discomfort affects their quality of life The subjects consisted of 315 female university students in Seoul City by convenience sampling from July 22 to August 3, 2010. The data was collected by structured questionnaire that included general characteristics, Menstrual Distress Questionnaire and Smith Kline Beecham 'Quality of Life' Scale. To analyze the data, used the following methods: frequency, mean, standard deviation, t-test, ANOVA, Pearson's correlation coefficients with the SPSS program 15.0. The Results as follows: The participants indicated water accumulation, the ache, negative emotion, conduct change and attention intensive obstacle order to show the women's menstrual discomforts and the quality of life was high competent feeling, body and mental stability, vitality and stability order. The correlation of the women's menstrual discomforts and the quality of life was r=-.605 and p=.000, the women's menstrual discomforts and the quality of life show considerably negative correlation. In the event that the women's menstrual discomforts was high comes to be low appeared the quality of life. As a result, this study showed that the women's menstrual discomforts and the quality of life have negative correlation, and discovered various factors which effect to the menstrual discomforts and the quality of life for the women's college students. Thus in the future, do a research for the middle school girls, the maiden and married women, and the repetition research of the women's college student is necessary. The nursing arbitration program development for relaxation of the women's menstrual discomforts is necessary as well.
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