• 폴리머
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• 제29권2호
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• pp.135-139
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• 2005

본 연구에서는 세포 독성을 감소시키고 in vivo에서 표적화를 향상시키기 위하여 methoxypoly(ethylene glycol) (mPEG) 및 folate가 도입된 poly(ethylene imine)(PEI)를 합성하였다. mPEG 말단에 카복실 그룹을 도입하여 EDC/NHS에 의해 활성화시킨 후 PEI의 아민기와의 반응으로 mPEG-PEI를 합성하였다. Folate의 카복실 그룹을 EDC/NHS으로 활성화시킨 후 mPEG-PEI의 아민기와의 반응으로 PEG-folate-graft-PEI를 합성하였다. 합성된 고분자의 화학적 구조는 $^1H-NMR$과 H-n을 이용하여 확인하였다. 합성된 고분자와 DNA가 정전기적 인력에 의해 완전한 복합체의 형성을 확인하기 위해 여러 가지 N/P charge 비율로 agarose gel 전기영동 및 형광을 측정하였고, 2 이상의 N/P charge 비율에서 완전한 복합체를 형성함을 확인하였다. 복합체의 크기는 광산란장치 및 AFM으로부터 $100\~300nm$임을 관찰하였다. 이러한 결과들로부터 합성된 양이온성 고분자와 DNA가 복합체를 잘 이루어지는 것을 관찰하였고 유전자 전달체로서의 가능성을 평가하고자 하였다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권4호
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• pp.309-314
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• 2005

We performed a basic experiment for the rapid, on-line, real-time measurement of hepatitis B surface antigen using a surface plasmon resonance biosensor. We immobilized anti­HBsAg (hepatitis B surface antigen) polyclonal antibody, as a ligand, to the dextran layer on a CM5 chip surface that had previously been activated by N-hydroxysuccinimide. A sample solution containing HBsAg was fed through a microfluidic channel, and the reflecting angle change due to the mass increase from the binding was detected. The binding characteristics between HBsAg and its polyclonal antibody followed the typical monolayer adsorption isotherm. When the entire immobilized antibody had interacted, no additional, non-specific binding occurred, suggesting the immunoreaction was very specific. The bound antigen per unit mass of the antibody was independent of the immobilized ligand density. No significant steric hindrance was observed at an immobilization density of approximately $17.6 ng/mm^2$. The relationship between the HBsAg concentration in the sample solution and the antigen bound to the ligand was linear up to ca. $40{\mu}g$/mL. This linearity was much higher than that of the ELISA method. It appeared the anti­gen-antibody binding increased as the immobilized ligand density increased. In summary, this study showed the potential of this SPR biosensor-based method as a rapid, simple and multi­sample on-line assay. Once properly validated, it may serve as a more efficient method for HBsAg quantification for replacing the ELISA.

• Asian-Australasian Journal of Animal Sciences
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• 제15권3호
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• pp.410-415
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• 2002

Adrenal glucocorticoids, secreted by the stimulus of adrenocorticotropic hormone (ACTH), are catabolic hormones in the pig. The present study was conducted to find whether active immunization against ACTH would suppress cortisol secretion accompanied by an increased growth rate in growing-finishing barrows. ACTH was conjugated to keyhole limpet hemocyanin or human histone using glutaraldehyde or 3-maleimidobenzoic acid N-hydroxysuccinimide, under a 2 (ACTH vs no hapten)${\times}$2 (carrier)${\times}$2 (crosslinker) factorial arrangement of treatments. Cross-bred barrows weighing approximately 25 kg were injected with an ACTHcarrier or carrier only conjugate every 4th wk and slaughtered at approximately 110 kg body weight. Antibodies against ACTH were detected in serum, as determined by $[^{125}I]$ACTH-binding activity, in most animals immunized against the ACTH conjugate, but not in carrier only-injected animals, except for the animals which had received the hapten conjugated to histone via glutaraldehyde. The $[^{125}I]$ACTH-binding activity of serum increased after the second booster injection, but overall ACTH antibody titer was very low. Main effect was not detected not only for the carrier and crosslinker but for the hapten in serum cortisol concentration, ADG, loin muscle area, backfat thickness and longissimus muscle composition including fat and protein. In addition, bound $[^{125}I]$ACTH percentage had no relation to cortisol concentration or to any of the above growth-related variables. Results suggest that ACTH or its conjugates used in the present study were not immunogenically potent enough to affect the glucocorticoid secretion and thus the growth of the immunized pigs.

• Journal of Microbiology and Biotechnology
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• 제27권8호
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• pp.1483-1490
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• 2017

In this study, silver nanoparticles (AgNPs) were synthesized by the citrate reduction process and, with the assistance of n-hydroxysuccinimide and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, were successfully loaded with the macromolecular drug vancomycin (VAM) to form AgNP-VAM bioconjugates. The synthesized AgNPs, VAM, and AgNP-VAM conjugate were characterized by UV-visible spectroscopy, zeta potential analysis, confocal microscopy, and transmission electron microscopy. The effect of loading VAM onto AgNPs was investigated by testing the internalization of the bioconjugate into Mycobacterium smegmatis. After treatment with the AgNP-VAM conjugate, the bacterial cells showed a significant decrease in UV absorption, indicating that loading of the VAM on AgNPs had vastly improved the drug's internalization compared with that of AgNPs. All the experimental assessments showed that, compared with free AgNPs and VAM, enhanced internalization had been successfully achieved with the AgNP-VAM conjugate, thus leading to significantly better delivery of the macromolecular drug into the M. smegmatis cell. The current research provides a new potential drug delivery system for the treatment of mycobacterial infections.

• Journal of Pharmaceutical Investigation
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• 제41권3호
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• pp.173-178
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• 2011

Injectable chitosan hydrogels have attracted great potential due to sustained-release property and safety. Here, palmityl-acylated glycol chitosan (Pal-GC) was used to generate physically cross-linked hydrogels by virtue of hydrophobic attraction of linear fatty carbons. Glycol chitosan was chemically modified with N-hydroxysuccinimide-activated palmitic acid in dimethylsulfoxide (DMSO) containing dimethylaminopyridine. Through a series of preparation steps of (i) dialysis with DMSO, (ii) addition of palmityl-acylated exendin-4 (Ex4-C16), and (iii) dialysis with water, Pal-GC was self-assembled to form physically cross-linked hydrogels entrapped with Ex4-C16. The Pal-GC derivative was analyzed by using 1H NMR, and the surface morphology of Pal-GC hydrogels formed was examined by scanning electron microscopy. Also, the hypoglycemic effect induced by Pal-GC hydrogels containing Ex4-C16 (250 nmol/kg) was evaluated in non-fasted type 2 diabetic db/db mice and compared with GC hydrogels containing native Ex4 at the same dose. Results showed that palmityl group was successfully conjugated with the amines of glycol chitosan, and that Pal-GC efficiently generated the hydrogels formation. Moreover, Pal-GC hydrogels containing Ex4-C16 was found to greatly prolong the hypoglycemia duration (~ 4 days). This was due to the dual-functions of the palmityl groups present in both GC and exendin-4 such as hydrophobic attraction and plasma albumin-binding. We consider this new type of self-assembled GC hydrogels loaded with Ex4-C16 would be a promising long-acting sustained-release system with anti-diabetic property.

• KSBB Journal
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• 제17권1호
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• pp.20-25
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• 2002

We performed a basic experiment for rapid, on-line, real-time measurement of HBsAg by using a surface plasmon resonance biosensor to quantify the recognition and interaction of biomolecules. We immobilized the anti-HBsAg polyclonal antibody to the dextran layer on a CM5 chip surface which was pre-activated by N-hydroxysuccinimide for amine coupling. The binding of the HBsAg to the immobilized antibody was measured by the mass increase detected by the change in the SPR signal. The binding characteristics between HBsAg and its antibody followed typical monolayer adsorption isotherm. When the entire immobilized antibody was interacted, there was no additional, non-specific binding observed, which suggested the biointeraction was very specific as expected and independent of the ligand density. No significant steric hindrance was observed at 17.6 nm/$mm^2$ immobilization density. The relationship between the HBsAg concentration in the sample solution and the antigen bound to the chip surface was linear up to ca. $40\mu\textrm{g}$/mL, which is much wider than that of the ELISA method. It appeared the antigen-antibody binding was increased as the immobilized ligand density increased, but verification is warranted. This study showed the potential of this biosensor-based method as a rapid, simple, multi-sample, on-line assay. Once properly validated, it can serve as a more powerful method for HBsAg quantification replacing the current ELISA method.

• Journal of Radiation Protection and Research
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• 제44권1호
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• pp.15-25
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• 2019

Background: Psammaplin A (PsA) is a radiosensitizer whereas its clinical application is hampered by poor bioavailability. This study aimed to synthesize novel radiosensitizers using PsA as the lead compound. Materials and Methods: Eight homodimeric disulfides were synthesized from corresponding acid and cystamine dihydrochloride in N-hydroxysuccinimide and dicyclohexylcarbodiimide coupling conditions. One monomeric thiol analog was obtained by reduction of homodimeric disulfide with dithiothreitol. Clonogenic assay was used to measure cell survival after irradiation and drug treatment in human lung cancer (A549) and glioblastoma (U373MG) cells. Results and Discussion: Using the PsA backbone, nine compounds were synthesized. Eight compounds showed variable cytotoxicity with 50% inhibitory concentrations ranging $16.14{\mu}M$ to $150.10{\mu}M$ (A549), and $13.25{\mu}M$ to $50.15{\mu}M$ (U373MG). Four and six compounds radiosensitized A549 and U373MG cells, respectively. Two compounds that radiosensitized both cell lines were tested for its inhibitory effects on DNMT1. One of them was shown to significantly inhibit DNMT1 activity. Conclusion: Novel compounds with radiosensitizing activity were synthesized. These compounds have a great potential to serve as a basis for the development of future radiosensitizers. Further investigation is warranted for their clinical application.

• Journal of Industrial and Engineering Chemistry
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• 제67권
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• pp.388-395
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• 2018

Significant efforts have been applied toward fabricating three-dimensional (3D) scaffolds using 3D-bioprinting tissue engineering techniques. Gelatin has been used in 3D-bioprinting to produce designed 3D scaffolds; however, gelatin has a poor printability and is not useful for fabricating desired 3D scaffolds using 3D-bioprinting. In this study, we fabricated pore size controlled 3D gelatin scaffolds with two step 3D-bioprinting approach: a low-temperature ($-10^{\circ}C$) freezing step and a crosslinking process. The scaffold was crosslinked with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). The pore sizes of the produced 3D gelatin scaffolds were approximately 30% smaller than the sizes of the designed pore sizes. The surface morphologies and pore sizes of the 3D gelatin scaffolds were confirmed and measured using scanning electron microscopy (SEM). Human dermal fibroblasts (HDFs) were cultured on a 3D gelatin scaffold to evaluate the effect of the 3D gelatin scaffold pore size on the cell proliferation. After 14 days of culture, HDFs proliferation throughout the 3D gelatin scaffolds prepared with more than $580{\mu}m$ pore size was approximately 14% higher than proliferation throughout the 3D gelatin scaffold prepared with a $435{\mu}m$ pore size. These results suggested that control over the 3D gelatin scaffold pore size is important for tissue engineering scaffolds.

• 한국수산과학회지
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• 제33권1호
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• pp.55-59
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• 2000

무지개 송어의 뇌하수체 및 배양액에 존재하는 GTH II 농도를 측정하기 위해 Avidin- Biotin complex를 이용한 sandwich EIA 계을 개발했다. Protein A sepharose affinity chromatography을 통해서 얻어진 연어 GTH II의 rabbit IgG에 biotinylation시킨 것 (Biotin-salmon GTH II rabbit IgG)을 제2 항체로 사용하였고, Non-Biotin salmon GTH II rabbit IgG는 단지 protein A sepharose affinity chromatography에서 얻어진 IgG를 제 1 항체로 사용하였다. EIA는 sandwich법에 의해서 이루어졌으며, 효소반응 기질로는 TMB(3,3'5,5-tetramethylbenzidine)를 이용했으며, 반응후 450 nm의 흡광도에서 automatic microplate reader로 측정하였다. 그 결과, $0.12\;{\~}\;125\;ng/ml$의 범위에서 용량반응곡선을 얻었으며, 측정감도 (최소 검출량)는 거의 0.58 ng/ml 정도 였다. 그리고 뇌하수체 추출물 및 배양액 각각의 희석곡선은 GTH II 표존곡선과 일치 하였다. 또한 이러한 GTH II의 표준곡선는 뇌하수체내 다른 peptide hormone와는 교차반응을 거의 나타내지 않았다. Testosterone을 처리한 미성숙 무지개 송어의 뇌하수체 세포배양계를 이용하여 sGnRH에 의한 GTH II 분비량을 본 sandwich EIA계와 RIA계를 비교 조사한 결과, 거의 같은 분비량을 나타냈을 뿐만아니라 같은 분비 pattern을 나타냈다. 이러한 결과로부터 본 sandwich법 EIA계에 의해서 연어과 어류의 뇌하수체 추출물 및 뇌하수체 배양액 중의 GTH II 함량 및 분비량을 측정하는데 있어서 안정된 assay계라고 생각되어진다.

• 폴리머
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• 제30권4호
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• pp.279-285
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• 2006

유전자 치료에 있어서 안전성의 장점을 지니고 있는 비바이러스성 전달체에 대한 관심도가 높아져가고 있다. 비바이러스성 전달체 중, 양이온성 리포좀이나 합성 유전자 전달체는 in vitro계에서 효율적인 DNA 전달체이지만, 낮은 생체적합성으로 인하여 in vivo 계에서의 응용성은 크게 뒤떨어지고 있다. 한편, 천연 양이온성 다당류인 키토산은 낮은 독성과 강한 양전하를 띠고 있어 유전자 전달 시스템 (gene delivery system)에 있어 아주 기대되는 전달체이다. 본 연구에서는 저분자량 수용성 키토산 (low molecular water-soluble chitosan ; LMWSC)을 이용하여 동맥 벽 세포를 표적할 수 있는 표적성 유전자 전달체를 합성하였다. 상대 점도와 Kina 적정법을 이용하여 LMWSC의 점도 평균 분자량 $(M_W)$과 탈아세틸화도 (degree of de acetylation ; DDA)를 측정하였고 구조는 FTIR, $^1H-NMR$, 그리고 $^{13}C-NMR$을 통하여 분석하였다. 동맥 벽을 표적하기 위한 유전자 전달체로서 pegylated LMWSC 의 말단에 특이성 세포 표적 펩타이드인 artery wall binding peptide (AWBP)를 결합시킴으로써 AWBP-PEG-g-LMWSC을 합성하였고 FTIR, $^1H-NMR$. zeta potentiometer. 그리고 atomic force microscopy (AFM)을 이용하여 분석하였다.