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Alpha - hCG 측정을 위한 섬광 근접 측정법 ( Scintillation Proximity Assay ) 에 관한 연구
최태현(Tae Hyun Choi),최창운(Chang Woon Choi),임상무(Sang Moo Lim),정위섭(Wee Sup Chung),임수정(Soo Jeong Lim),이수진(Su Jin Lee),이태섭(Tae Sup Lee),오옥두(Ok Doo Awh),(Kwang Sun Woo) 대한핵의학회 2002 핵의학 분자영상 Vol.36 No.2
목적: 섬광 근접측정법은 항원 항체 반응 후 결합 분획과 유리분획을 분리하는 광정이 필요없다. 이러한 원리를 검체 내 hCG와 항 α hCG 항체간의 항원 항체 반응에 적용하고자 한다. 대상 및 방법: 항 α hCG 항체를 biotin과 결합시켜 SPA bead에 부착된 streptavidin과 부착 가능하게 만들었다. 이 측정법은 항 α hCG 항체가 부착된 SPA beads에 대해 혈청내 hCG와 표지항원인 [^125I]hCG간의 경쟁 반응을 기본 원리로 이용하였다. Biotin표지 항 α hCG항체를 [^125I]hCG 100㎕와 표준용액이나 환자 혈청 200㎕이 들어있는 실온에서 20분 방치하였다. 그리고 streptavidin이 붙은 SPA beads 20㎕를 바이알에 넣고 10분 더 방치한다. 환자 혈청의 수치를 표준 응답곡선을 통해 계산하였다. 결과: SPA측정법에 사용되는 방사성 핵종의 방사능 양에 따라 반응용액 속에서 SPA bead와 자유 방사성 핵종에 의한 배후 방사능이 측정값에 영향이 없음을 확인하였다. SPA 방법을 응용한 측정에서 적합한 표준 응답곡선을 얻었고, 실제 환자혈청에서의 hCG농도를 결정할 수 있었다. 결론: 이 실험을 통해 SPA 방법을 이용한 측정법이 임상진단에 유용하게 사용될 수 있을 것으로 사료된다. Purpose: Scintillation Proximity Assay (SPA) does not require the physical separation of receptor bound form from free form. SPA was applied to the study of interaction of human chorionic gonadotrpin (hCG) and anti-α hCG in serum. Materials and methods: Anti-α hCG was biotinylated for the binding to streptavidin. The assay was based on the simple competitive binding method between [^125l]hCG and the hCG in sample serum, with anti-α hCG-coated beads. Aliquots of biotinylated anti-α hCG were dispensed into scintillation vials containing 100㎕ [^125ㅣ]hCG and 200㎕ of either a standard concentration of hCG for preparation of standard curve or unknown sample, and oncubated for 20 min. at room temperature. Then 20㎕ streptavidin-coated beads were added to vials. and finaliy incubated for 10 min at room temperature. Values for unkncown samples were then calculated from the standard curve. Results: Optimal backfround counts were certificated using varied radioactivity of radionuclides. Appropriate standard curve was obtained from SPA method successively, and the concentration of hCG from unknown serum was determined by standard curve. The result from SPA assay was similar to that of RIA. Conclusion: This observation confims that SPA method could be useful for cilnical diafnosis.(Korean J Nucl Med 2002;36;133-139)
간암 동물 모델에서 2`-fluoro-2`-deoxy-1-β-D-arabinofuranosyl-5-[124I]iodo-uracil ([124I]FIAU) 소동물 PET 영상 연구
채민정 ( Mim Jeong Chae ),이태섭 ( Tae Sup Lee ),김준엽 ( June Youp Kim ),우광선 ( Gwang Sun Woo ),정위섭 ( Wee Sup Jumg ),전권수 ( Kwon Soo Chun ),김재홍 ( Jae Hong Kim ),이지섭 ( Ji Sup Lee ),류진숙 ( Jin Sook Ryu ),천기정 ( Gi 대한핵의학회 2008 핵의학 분자영상 Vol.42 No.3
송인호 ( In Ho Song ),이태섭 ( Tae Sup Lee ),강주현 ( Joo Hyun Kang ),이용진 ( Yong Jin Lee ),김광일 ( Kwang Il Kim ),안광일 ( Gwang Il An ),정위섭 ( Wee Sup Chung ),천기정 ( Gi Jeong Cheon ),최창운 ( Chang Woon Choi ),임상무 ( Sa 대한핵의학회 2009 핵의학 분자영상 Vol.43 No.5
목적: Hydrodynamic-based procedure는 손쉽고 간편한 비바이러스성 유전자 전달 방법으로 특히 간특이적으로 발현하는 특징을 가진다. 단순 헤르페스 바이러스 제 1 형 티미딘 키나제(herpes simplex virus type 1 thymidine kinase, HSV1-tk)와 다양한 기질을 이용한 비침습적 HSV1-tk 유전자 영상시스템이 널리 연구되어왔다. 본 연구에서는 HSV1-tk 유전자를 hydrodynamic-based procedure를 이용하여 전달한 후, HSV1-tk의 보고 기질로 알려진 5-(2-iodovinyl)-2`-deoxyuridine (IVDU)을 이용하여 간 특이적인 HSV1-tk 유전자 발현 영상을 획득하고자 하였다. 대상 및 방법: HSV1-tk 유전자와 녹색형광유전자를 가진 각 플라스미드 벡터를 마우스에 hydrodynaminc injection을 통해 전달하고, 24 시간 뒤 유전자의 발현을 확인하기 위해 RT-PCR, 생체형광영상, 핵의학영상, 전신자가방사영상 그리고 생체분포를 시행하였다. 결과: 각 플라스미드 벡터를 전달한 간으로부터 추출한 전체 RNA를 이용하여 RT-PCR을 수행한 결과, 각각 HSV1-tk 유전자와 녹색형광단백 유전자의 특이적인 밴드를 관찰할 수 있었다. 생체 분포 결과, pHSV1-tk 벡터를 전달한 마우스의 간에서 특이적인 [123I]IVDU의 섭취를 보였다. 생체형광영상에서는 pEGFP-N1 벡터를 전달한 마우스의 간에서는 유의한 형광신호를 나타내었다. 전신자가방사영상과 감마카메라 영상에서 pHSV1-tk 벡터를 전달한 마우스의 간에서 방사표지 IVDU가 국소적으로 집적되는 것을 확인하였다. 결론: 본 연구에서 hydrodynamic-based procedure는 간특이적으로 플라스미드 DNA를 전달하는데 효과적이며 전달된 유전자의 발현을 분자영상학적인 방법으로 확인하였다. 따라서 Hydrodynamic injection을 통해 HSV1-tk 유전자와 목적 유전자의 공동발현은 방사표지 IVDU에 의해 목적 유전자의 발현을 정량평가하는데 유용할 것으로 기대된다. Purpose: Hydrodynamic-based procedure is a simple and effective gene delivery method to lead a high gene expression in liver tissue. Non-invasive imaging reporter gene system has been used widely with herpes simplex virus type 1 thymidine kinase (HSV1-tk) and its various substrates. In the present study, we investigated to image the expression of HSV1-tk gene with 5-(2-iodovinyl)-2`-deoxyuridine (IVDU) in mouse liver by the hydrodynamic-based procedure. Materials and Methods: HSV1-tk or enhanced green fluorescence protein (EGFP) encoded plasmid DNA was transferred into the mouse liver by hydrodynaminc injection. At 24 h post-injection, RT-PCR, biodistribution, fluorescence imaging, nuclear imaging and digital wholebody autoradiography (DWBA) were performed to confirm transferred gene expression. Results: In RT-PCR assay using mRNA from the mouse liver, specific bands of HSV1-tk and EGFP gene were observed in HSV1-tk and EGFP expressing plasmid injected mouse, respectively. Higher uptake of radiolabeled IVDU was exhibited in liver of HSV1-tk gene transferred mouse by biodistribution study. In fluorescence imaging, the liver showed specific fluorescence signal in EGFP gene transferred mouse. Gamma-camera image and DWBA results showed that radiolabeled IVDU was accumulated in the liver of HSV1-tk gene transferred mouse. Conclusion: In this study, hydrodynamic-based procedure was effective in liver-specific gene delivery and it could be quantified with molecular imaging methods. Therefore, co-expression of HSV1-tk reporter gene and target gene by hydrodynamic-based procedure is expected to be a useful method for the evaluation of the target gene expression level with radiolabeled IVDU. (Nucl Med Mol Imaging 2009;43(5):468-477)
3′-[(18)F]Fluoro-3′-deoxythymidine의 합성과 9L glioma 세포를 이식한 래트에서의 체내동태에 관한 연구
심아영 ( Ah Young Shim ),문병석 ( Byung Seok Moon ),이태섭 ( Tae Sup Lee ),이교철 ( Kyo Chul Lee ),안광일 ( Gwang II An ),양승대 ( Seung Dae Yang ),유국현 ( Kook Hyun Yu ),천기정 ( Gi Jeong Cheon ),최창운 ( Chang Woon Choi ),임상 대한핵의학회 2006 핵의학 분자영상 Vol.40 No.5
지방육종형성 동물모델에서 123I-15-(p-iodophenyl)-3-R , S-methylpentadecanoic acid ( BMIPP ) 의 생체분포와 생체영상
최창운(Chang Woon Choi),임상무(Sang Moo Lim),이태섭(Tae Sup Lee),서용섭(Yong Sup Suh),우광선,정위섭(Wee Sup Chung),임수정(Soo Jung Lim),오옥두(Ok Doo Awh) 대한핵의학회 2001 핵의학 분자영상 Vol.35 No.5
N/A Purpose: 123I-labeled fatty acids have been used in the evaluation of regional myocardial energy metabolism. This study aimed to evaluate the usefulness of 123I-BMIPP as a liposarcorna-imaging agent. Materials and Methods: We compared in vitro uptakes between liposarcoma(SW872) and glioma(9L) cell lines, and examined biodistribution and in vivo images of 123I-BMIPP in liposarcoma-bearing nude mice. Cold-BMIPP was labeled with 123I using Cu2+ as catalyst. After purification by Sep-pak, radiochemical purity was determined by TLC. We compared cellular uptake between glioma and liposarcoma after incubation of 5, 10, 15, 30, 60, 120, and 180 mins with culture medium containing I-BMIPP. The difference in biodistribution was determined between non-feeding (water only) group for 18 hr and feeding group in normal mice (n=6/group) at 0.5, 2, and 24 hr. In liposarcoma-bearing nude mice model, liposarcoma, SW872, cell lines were injected subcutaneously into the left thigh of nude mice. The biodistribution of 123I-BMIPP was evaluated at 0.5, 2, and 24 hr (n=5 / group) and in vivo image of 123I-BMIPP was obtained with gamma camera at 2 and 24 hr in liposarcorna-bearing nude mice. Results: Radiolabeling yield and radiochemical purity were 95% and above 99%, respectively. SW872 cell line showed more increased uptake than 9L with 1.5 times at 180 mins. The clearance of 'I-BMIPP in various tissues was more delayed in the non-feeding group than in the feeding group, especially at delayed time (24 hr) in normal mice, and the major excreting organ was the gastrointestinal tract. In liposarcoma-bearing nude mice, tumor/blood ratio of 123I-BMIPP was 0.94, 0.75, and 1.38 and tumor/muscle ratio was 0.66, 1.53, and 1.11 at 0.5, 2, and 24hr, respectively. 123I-BMIPP was selectively localized in liposarcoma at 24 hr image. Conclusions: These results suggest that 123I-BMIPP can be used as a liposarcoma-imaging agent. (Korean J Nucl Med 200135:324-333)
Taxol 방사면역측정을 위한 125I-Iodotyraminehemisuccinyltaxol ( 125ITHT ) 의 제조
최창운(Chang Woon Choi),임상무(Sang Moo Lim),오옥두(Ok Doo Awh),이태섭(Tae Sup Lee),최태현(Tae Hyun Choi),김현석(Hyun Suk Kim),홍준표(Jun Pyo Hong),이은숙(Eun Sook Lee) 대한핵의학회 2002 핵의학 분자영상 Vol.36 No.2
목적: Taxol(Paclitaxel)은 난소암과 유방암의 치료제로 사용되고 있으며, 치료시 적절한 체내 혈중농도를 유지함으로서 치료효과를 극대화하기 위해서는 taxol의 혈중농도를 측정할 수 있는 방사면역측정시스템에 표지항원으로 사용할 수 있는 taxol 유도체의 방사성표지화합물을 합성하고, 이를 이용하여 방사면역측정법을 시행할수 있는지의 여부를 확인하고자 하였다. 대상 및 방법: Taxol과 succinic anhydride를 무수 pyridine을 용매로 하여 반응시켜 hemisuccinyltaxol을 합성하고, 합성된 hemisuccinyltaxol과 tyramine을 isobutyl-chloroformate를 coupling agent로 사용하여 tyraminehemisuccinyltaxol을 합성하고 HPLC로 분리정제 하였다. 산화제인 Chloramine-T(5.25 ㎎/ml, 10㎕)를 사용하여 tyraminehemisuccinyltaxol(4 ㎎/ml, 30㎕)에 ^125l(l mCi)를 방사성요오드화하고 HPLC를 이용하여 표지수율을 산정하였다. 정제된 tyraminehenisuccinyltaxol과 ^125l-iodotyraminehemisuccinyltaxol을 80% acetonitrile 수용액에 녹여 4℃와 37℃로 보관하면서, 각 시간대별로 화학적 순도와 방사화학적 순도를 결정하여 그 안정도를 HPLC를 이용하여 확인하였다. [^125l]Iodotyraminehemisuccinyltaxol를 방사성표지항원으로 사용하여 taxol에 대한 단클론항체(3G5A7)의 역가를 검정하였으며, 0∼100nM 농도범위에서 taxol 농도의 증가에 따른 표준투여응답곡선을 작성하였다. 결과: Hemisuccinyltaxol은 79.9%의 수율로 합성되었으며, tyraminehemisuccinyltaxol의 합성수율은 19.5%였다. ^125l-iodotyraminehemisuccinyltaxol의 표지수율은 93%이었다. tyraminehemisuccinyltaxol은 7일까지도 96.5% 이상의 순도를 보여 비교적 안정함을 확인하였으며, ^125l-iodotyraminehemisuccinyltaxol은 3일까지는 93.4% 이상으로 안정하였고 또한 7일 경과시에는 86.1% 이상의 순도를 보였다. taxol에 대한 단클론항체(3G5A7)의 역가를 검정하여 1:256의 역가를 나타냄을 확인하였으며, taxol 농도에 따른 표준투여응답곡선은 taxol과 방사표지 taxol 유도체간에 경쟁적으로 사용되어 직선성(R2=0.971)을 나타내었다. 결론: taxol의 경쟁적 방사면역측정법의 방사성 추적자로서 방사성표지 taxol 유도체인 ^125l-iodotyraminehemisuccinyltaxol을 이용한 방법이 유용함을 확인하였다. Purpose: Taxol(Paclitaxel), an antineoplastic agent, has been used in the treatment of ovarian and breast cancers. The determination of optimal Taxol concentrations in human serum was required for enhancing therapeutic effect and maintaining the appropriate Taxol level in blood. This study was aimed to synthesize radiolabeled Taxol derivatives as radiotracer in competitive radioimmunoassay for monitoring Taxol concentrations in blood and to determine the usefulness of its derivatives. Materials and Methods: Hemisuccinyltaxol(HT) was synthesized by esterification of Taxol with succinic anhydride. Tyraminehemisuccinyltaxol(THT) was synthesized by coupling of HT with tyramine using isobutylchloroformate as coupling agent and purified by HPLC. By using chloramine-T(5.25 ㎎/ml, 10㎕) as oxidant agent, THT(4 ㎎/ml, 30㎕) was labeled with ^125I(37 MBq, 1 mCi). To estimate the stability of purified THT, ^125I-iodotyraminehemisuccinyltaxol(^125ITHT) was dissolved in 80% acetonitrile aqueous solution, and the solution was incubated at 4℃ and 37℃ for 7 days. At various time intervals, the stability of THT and ^125ITHT was monitored. The tiler of Taxol monoclonal antibody, 3G5A7, was determined by competitive radioimmunoassay using ^125ITHT as a labeled antigen. A standard dose-response curve was demonstated by Taxol competitive radioimmunoassay. Results: HT and THT were synthesized with 79.9% and 19.5% yield, respectively. The labeling yield of ^125ITHT was 93%. After 7 days, the chemical purity of THT was 96.5% at 4℃, and 97.5% at 37℃. After 3 days, ^125ITHT was stable with 94.7% at 4℃ and 93.4% at 37℃. After 7 days, radiochermical purity was diminished to 88.1% at 4℃ and 86.1% at 37℃. The titer of Taxol monoclonal antibody, 3G5A7, was determined to 1:256. A standard dose-response curve demonstated good colinearity (R^2=0.971) as Taxol concentration-dependent manner. Conclusion: Competitive radioimmunoassay using ^125I-iodotyraminehemsuccinytaxol as radiotracer could be used to monitor for concentration of Taxol in the human serum.