뇨 증 삼중수소 측정시 방부제 첨가 효과

오옥두 연세대학교 보건과학연구소 2001 보건과학논집 Vol.11 No.-

뇨중 삼중수소 측정

오옥두,송명재 연세대학교 보건과학연구소 1998 보건과학논집 Vol.8 No.-

디메틸설폭사이드 표지보조제를 사용한 인슈린의 방사성요드화

오옥두 연세대학교 보건과학연구소 1996 보건과학논집 Vol.6 No.-

방사면역측정용 T₃항체 고정화 연구

吳玉斗,白春善 연세대학교 보건과학연구소 1994 보건과학논집 Vol.4 No.-

T3-BSA,T4-BSA 접합체 제조 및 생성항체의 방사면역측정적 이용 연구

오옥두,김재록 대한핵의학회 1980 핵의학 분자영상 Vol.14 No.1

1) T₃-BSA 및 T₄-BSA는 Morpho CDI를 coupling agent로 사용하여 pH 5.5에서 12시간 정도 실온 하(10∼20℃)에 반응시키는 것이 효고적이며 특히 T₄의 경우는 DMF를 충분히 가해야 침전생성을 막을 수 있다. 2) 제조한 T₃-BSA 및 T₄-BSA의 접합 mole비율은 각각 8∼9:1 및 4∼5:1이었다. 이들을 사용하여 얻은 항혈청의 RIA용 적정 희석비율은 각각 1.5×104:1 및 2×103:1이었으며 교차반응성은 T₄항체가 T3항체에 비해 7∼8배 컸다. 따라서 T₄항체가가 낮게 얻어지는 이유는 T₄-BSA의 접합비율이 낮다는 사실 이외에 T₄-BSA접합체 제조, 정제, 면역등 일련의 과정에서 T₄-BSA의 T₃- BSA로의 분해에 기인한다고본다. 3) T₃나 T₄를 ester화 하지 않은채로 만든 접합체를 써서 얻은 항혈청으로도 RIA용 적정 항혈청 희석비율율에서의 교차반응영향은 무시됨으로 T₃, T₄ RIA가 가능하였다. T₃-BSA, T₄-BSA conjugates were prepared and identified spectrophotometrically. The lambda-max on the conjugates was just coincided with that of BSA, but the molar extinction coefficients of the conjugates were generally larger than that of BSA itsalf. The molar ratios of T₃:BSA and T₄:BSA in the prepared conjugates were found to be 9:1 and 5:1, respectively. The titers of the T₃ antisera were generally higher (max. 1.5×104:1) than those of T₄ (max.2×103:1), and the average cross reactivity of the T3 antibody with T₄ was lower (0.45%) than that of T4 antibody with T₃(3∼4%). The results of the study indicate that the predominant cause of the lower titers and the lower specificity of the T₄ antisera comparing with those of T₃ is mainly due to the unstability of the T₄-BSA and consequent degradation of the conjugate to T₃-BSA during preparation, purification, and even during immunization. The lower molar ratio of T₄ to BSA in the preparation stage is also considered to be a minor factor. By measuring T₃, T₄ les in the reference control serum, it has been confirmed that the prepared antisera can sufficiently be utilized, respectively, in the established radioimmunoassay systems.

Taxol 유도체들의 생물학적 거동에 관한 연구

오옥두,유대웅,임상무 ( Ok Doo Awh,Dae Wung Yoo,Sang Moo Im ) 대한핵의학회 1997 핵의학 분자영상 Vol.31 No.4

This study was designed to prospect the 'In-labelled paclitaxel as tumor imaging agent. In order to provide a taxol molecule with a functional group which is able to chelate In-lll, taxol-DTPA conjugate and 2-hemisuccinyltaxol were synthesized by esterification of taxol at C-2 on C-13 carbon with DTPA anhydride and succinic anhydride, respectively. Synthesis yield of the taxol derivatives was 34% for taxol- DTPA and 80% for 2'-hemisuccinyltaxol. Cytotoxicity of the taxol derivatives were measured by MTT method toward cell lines HT29, B16, P388, and CT26. The cytotoxic activities of the taxol derivatives were maintained, although less active than taxol. Radiolabelling of the taxol derivatives were proceeded directly with InCh or indirectly with In-citrate(ligand-exchange method). The ligand-exchane methocl was not suitable because some precipitat:es appeared during the reaction. On the contrary, by direct radiolabelling methnd, we were able to obtain taxol DTPA--'In in 100% radiochemical yield. However, 2'-hemisuccinyltaxol was not labellecl by both methods. Yield and radiochemiral purity of the radiolabelled com- pound were determined by HPI.C, paper chromatography and instant thin layer chromatography. Taxol-DTPA-In was characterized to be hydrophilic by lipophi- licity test, and nearly non-adhesive to HT29, E316, P388, and CT26 by cell hinding affinity test. Binding affinity of the taxol-DTPA-'In complex to serum proteins was also examined by protein precipitation with 30% trichloroacetic acid. The results showed that 309o of the taxol-DTPA-'In complex binds with serum proteins.

T₃항체생산을 위한 T₃-BSA합텐 접합체 제조연구

吳玉斗,白春善 연세대학교 보건과학연구소 1993 보건과학논집 Vol.3 No.-

농양진단을 위한 IgG-188Re 표지화합물 제조

오옥두,임상무,최태현 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1997 Journal of biomedical laboratory sciences Vol.3 No.2

농양진단을 위한 방사성표지화합물의 제조에 관한 기초실험을 수행하였다. IgG를 2-mercaptoethanol로 환원하여 분자당 1.5개의 -SH가 유도된 IgG를 얻을 수 있었다. 이것을 188Re과 표지 반응시켜 99%의 높은 표지반응수율로 IgG-188Re을 얻었으며, 여기에 인 혈청을 안정제로 가해줌으로써 1시간까지 약 90%의 방사화학적 순도를 유지할 수 있었다. 포도상구균으로 유발한 농양이식 백서에서 IgG-188Re의 생체분포실험을 통해 농양의 진단이 가능한 것으로 확인하였다. 이상의 결과를 적용하면 여러 가지 단클론 항체의 188Re표지화합물 제조에 적용할 수 있을 것으로 기대되었다. IgG-188Re conjugate was prepared for the diagnosis of abscess. The IgG molecules reduced by 2-mercaptoethanol contained 1.5 free sulfhydryl groups per IgG molecule. The reduced IgG molecule was labelled with 188Re through chelate to 99% of labelling yield. The radiochemical purity of IgG-188 Re conjugate was maintained at 90% in the presence of human serum for 1 hour. The IgG-188Re was intravenously administered into staphylococcal abscess-bearing rats and their biodistribution was monitored at 4 and 24 hours post injection. The IgG-188Re conjugate was moderately localized in the abscess tissue. This result implies that the IgG-188Re conjugate can be a tool for abscess diagnosis. This technique can be applied for the preparation of various monoclonal antibody labelled with 188Re.

Taxol의 방사면역측정을 위한 I-125 표지화합물 합성

오옥두,최창운,금준섭,박용석,이양호,편웅범 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1997 Journal of biomedical laboratory sciences Vol.3 No.2

Taxol은 diterpenoid구조를 가진 항암제로서, 난소암과 유방암에 탁월한 효과를 보이지만 다른 항암제와 마찬가지로 독성을 가지고 있어 약물의 체내 혈중농도를 모니터링하는 것이 필요하다. 약물의 혈중농도를 모니터링하는 방법은 HPLC법, ELISA법, RIA법 등이 있으나, RIA법이 민감도 측면에서 또한 간편하다는 점에서 장점이 있다. 본 연구에서는 I-125 표지항원을 이용한 방사 면역측정법을 확립하기 위해 먼저 taxol유도체를 합성하였다. 먼저 taxol의 C-13 탄소의 곁가지에 위치한 C-2'부분의 hydroxy기를 succinic anhydride와 반응시켜 2'-hemisuccinyltaxol(Ⅰ)을 합성(반응수율:80%)하였다. 또한 tyramine을 125I로 표지하고 gel chromatography를 통해 정제된 [125I]iodotyramine(Ⅱ)(반응수율:58%)을 얻었다. (Ⅰ)과 (Ⅱ)를 반응시켜 2'-[125I]iodotyramine-hemisuccinyltaxol (Ⅲ) (반응수율:96%)을 얻어 125I 표지항원으로 사용하였다. Taxol에 대한 항체를 획득하기 위해서 (Ⅰ)을 BSA에 접합반응시켜 2'-hemisuccinyltaxol-BSA접합체를 합성하였으며, 이것을 토끼에 면역주사하여 anti-taxol serum을 얻었다. 이 항체에 대한 역가 검정실험에서 1:20의 희석비에서 B/F(%)가 약 40%를 보였다. 이와 같은 결과는 2'-[125I]iodotyramine-hemisuccinyltaxol을 표지항원으로 한 taxol의 방사면역측정 방법으로 혈청내 taxol의 농도측정이 가능함을 제시해 준다. Taxol, an anticancer drug that has diterpenoid conformation, has been used as an effctive chemotherapeutical agent in the treatment of breast and ovarian cancers. Because of its toxicity like other anticancer drugs, monitoring the taxol level in serum is important procedure during cancer therapy. The various monitoring methods using HPLC, ELISA, and RIA have been adopted, and RIA technique is known to be superior than other methods in trems of sensitivity and convenience. In this study, in order to develope taxol RIA system using 125I labelled antigen, first of all we synthesized taxol derivatives. 2'-hemisuccinyltaxol was obtained with about 80% yield by esterification of taxol at C-2' hydroxyl group on C-13 carbon with succinic anhydride. [125I]iodotyramine was prepared with 58% labelling yield by radioiodination of tyramine and purified by gel chromatography. 2'-[125I]iodotyramine-hemisuccinyltaxol, 125I labelled antigen for taxol RIA, was synthesized with 96% yield from conjugation of 2'-hemisuccinyltaxol and [125I] iodotyramine. Anti-taxol serum was produced from the rabbit immunized with 2'-hemisuccinyltaxol-BSA synthesized by 2'-hemisuccinyltaxol and BSA. The antiserum titer was determined by RIA using 2'-[125I]iodotyramine-hemisuccinyltaxol. The titer of 1:20 was obtained with about 40% of B/T. The results suggest that taxol RIA using 125I labelled antigen can be applied to monitor the taxol level in serum.

Methionine 이성질체들의 99mTc 착물 제조 연구

오옥두(Ok Doo Awh),장희순(Hee Soon Chang),이동선(Dong Sun Lee) 대한핵의학회 1992 핵의학 분자영상 Vol.26 No.1

N/A Tc-99m-Methionine complexes from enantiomeric and racemic methionines were prepared controlling reaction parameters such as pH and the concentration of stannous chloride. Some radiochromatographic systems were also examined to determine the labelling yields of Tc-99m complexes. The best resolutions of Tc-99m complexes were obtained at ITLC-SA developed with acetone and paper chromatography with n-butanol saturated with 0.3N HCI. In the former system, HR-Tc-99m and Tc-99m-methionine complex remained at origin, while 99mTcO4 moved with Rf value of 1.0. In latter process, HR-Tc-99m stayed at the origin, while 99mTcO4 and Tc-99m- methionine complexes moved with Rf value of 0.5. By combining of two chromatographic systems, the contents of three Tc-99m species were calculated easily. Tc-99m Labelling from enantiomeric and racemic methionines had little differences and the optimal condition was found at pH 9.00 and the molar ratio of methionine to stannous chloride od 24:1. The yields of Tc-99m complexes from D-, L-, and DL-methionines were 87.6%, 94.1%, and 97.9%, respectively. The results indicated that methionine containing relatively hydrophobic methylthio group (-SCH3) would be labelled with Tc-99m by stannous chloride method.