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Efficiency delivery of polyplex based on microneedle with enhancing immune response
김낙원,정지훈 한국공업화학회 2015 한국공업화학회 연구논문 초록집 Vol.2015 No.1
The approach of gene delivery with microneedle has been got many attentions in diverse studies. Although the outstanding capability of carrying DNA into body with microneedle, there is a problem on stability of DNA coated on microneedle surface when it is stored during plenty of times in room temperature. And the period of generating immune responses is important portion to verify efficiency vaccine system with microneedle. In this study, we developed the novel polypelxes delivery system based on microneedle which was accompanied with polypelxes and FMLP peptides. We could be sure that the polplexes were delivered stably and FMLP peptide was able to increase immune responses due to the ability of immune cell recruitment. Also, the microneedle injection showed more excellent induction of antigen and antibody expression than subcutaneous injection.
김낙원,고보성,김애선,정지훈 한국공업화학회 2015 한국공업화학회 연구논문 초록집 Vol.2015 No.0
The approach of gene delivery with microneedle has been got many attentions in diverse studies. Although the outstanding capability of carrying DNA into body with microneedle, there is a problem on stability of DNA coated on microneedle surface when it is stored during plenty of times in room temperature. And the period of generating immune responses is important portion to verify efficiency vaccine system with microneedle. In this study, we developed the novel polypelxes delivery system based on microneedle which was accompanied with polypelxes and fMLP peptides. We could be sure that the polplexes were delivered stably and fMLP peptide was able to increase immune responses due to the ability of immune cell recruitment. Also, the microneedle injection showed more excellent induction of antigen and antibody expression than subcutaneous injection.
Bacillus subtilis JK-56이 생산하는 chitinase isozyme의 정제와 특성 규명
김낙원,정영기,전홍기 부산대학교 유전공학연구소 2002 분자생물학 연구보 Vol.18 No.-
토양으로부터 chitinase를 생성하는 균주를 분리하여 동정한 결과 Bacillus subtilis로 판명되었으며, 분리한 균주를 Bacillus subtilis JK-56이라 명명하였다. B. subtilis JK-56의 chitinase 생산 최적 조건을 검토한 결과 1% chitin, 0.5% polypeptone, 0.1% KCl, 0.05% MnSO_4 · 4H_2O이며 초발 pH 7.0, 배양온도 37℃에서 가장 많은 효소를 생산하였다. 본 균주가 생산하는 chitinase를 정제하기 위해서 native-PAGE를 이용해 효소활성 band를 확인한 결과, 1개의 강한 활성 band와 2개의 약한 활성 band를 가지는 isozyme중 1개의 강한 활성 band를 정제하였고 정제된 효소를 Chi-56A라고 명명하였다. Chi-56A의 효소 특성에 관해서 실험한 결과 분자량은 약 53kDa, pl는 4.3으로 확인되었다. 본 효소는 65℃까지 상당히 안정하였으며 효소의 최대활성 온도도 65℃로 확인되는 등 열에 대해 상당히 안정한 효소로 확인되었다. Collidal chitin에 대한 정제효소 Chi-56A의 K_m값은 17.33g/L였다. 그리고 pH 6.0에서 최대의 활성을 나타내었고, 산성범위보다 알칼리범위에서 안정한 것으로 나타났다. 또한 Mn^2+ 존재 하에서 높은 활성을 나타내었고 Co^2+와 Mg^2+ 존재 하에서도 활성이 약간 증가한 반면에 Hg^2+ 존재 하에서는 상당한 저해를 받았다. Chito 올리고당에 대한 분해 산물을 HPLC로 확인해 본 결과 짝수개의 올리고당의 분해산물은 (GlcNAc)_2만을 생산하였고 홀수개의 올리고당에 대해서는 GlcNAc와 (GlcNAc)_2를 생산하는 것으로 비환원성 말단으로부터 이당체인 diacetyl chitobiose ((GlcNAc)_2)를 생산하는 exo형 chitinase로 추정된다. Chitin, a β-1,4 polymer of N-acetyl-D-glucosamine, is one of the most abundant organic compounds in nature. Chitinase (EC 3.2.1.14) is an enzyme that degrades chitin to chito-oligosaccharides, diacetyl chitobiose and N-acetyl-D-glucosamine. An extracellular chitinase-producing bacterial strain was isolated from soil and named to as Bacillus subtilis JK-56. Optimum culture condition of B. subtilis JK-56 for the production of chitinase was 1% chitin, 0.5% polypepton, 0.1% KCl, 0.05% MnSO_4 · 4H_2O, 37℃, initial pH 7.0 and 40 hour culture time. When B. subtilis JK-56 was grown in the optimum medium, one major active band and two minor active bands were detected by native-PAGE and active staining of the gel. Among them, the major band was purified from the culture supernatant by 70% ammonium sulfate precipitation and native-PAGE with BIO-RAD Model 491 Prep-Cell and named as Chi-56A. Its molecular weight was estimated to be 53kDa monomer and the isoelectric point (pl) was pH 4.3. The pH and temperature for the optimum activity of Chi-56A were pH 6.0 and 65℃, respectively. Chi-56A was stable up to 65℃ and in alkaline region. Its K_m value for colloidal chitin was 17.33g/L. HPLC analysis of the reaction products confirmed that Chi-56A was an exo type chitinase.
김낙원,이민상,이정은,조용우,정지훈 한국공업화학회 2014 한국공업화학회 연구논문 초록집 Vol.2014 No.1
The approach of gene delivery with microneedles has recently been withdrawn much attention. Despite superior capability of microneedles delivering DNA into the dermal tissues, release and cellular uptake of the released DNA after microneedle-based administration have not been clearly studied. In this study, we fabricated polyplex-releasing multilayered microneedles additionally coated with microspheres containing a chemo-attractant for recruiting professional antigen presenting cells such as macrophages. The microspheres can be released from the needle body and localized in the dermis area where chemo-attractant slowly released. The ability of the microneedle-based cutaneous gene delivery system for attracting antigen presenting cells and improving DNA vaccination was tested.
파장가변형 흡수분광법을 이용한 질산암모늄 생성 과정에서의 NO와 NH₃ 농도 실시간 변동 측정
정낙원(Nakwon Jeong),소성현(Sunghyun So),김대해(Daehae Kim),송순호(Soonho Song),이창엽(Changyeop Lee),박대근(Dae Geun Park) 한국에너지기후변화학회 2020 에너지기후변화학회지 Vol.15 No.2
As rapidly growing across industries, particulate matters, which become a social issues in Korea, are caused by using Fossil fuels and expanding transportation. By atmospheric chemical reactions, primary air pollutants convert into secondary air pollutants. These pollutants decrease visibility and cause environmental problems such as acid rain and harmful effects on human health. Gaseous NO and NH₃ are significant precursor of secondary air pollutants, it is important to measure concentration of these gases. TDLAS, which is an optical measurement technology, has several advantages such as non-invasive, sensitive, fast response and robustness against various forms of noise, allow measuring the transient analysis of particulate matters. In this paper, the reaction relationship with NO, NH₃, H₂O and O₃ was analyzed by measuring the change in gaseous concentration during the conversion of nitrogen oxides to ammonium nitrate using TDLAS.