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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.
Convenient Preparation of Tumor - specific Immunoliposomes Containing Doxorubicin
Park, Yong Serk,Nam, Sang Min,Cho, Jang Eun,Son, Byoung Soo 생화학분자생물학회 1999 BMB Reports Vol.31 No.1
Two innovative methods to prepare target-sensitive immunoliposomes containing doxorubicin by coupling monoclonal antibodies (mAb DH2, SH1) specific to cancer cell surface antigens (G_(M3), Le^X) have been developed and are described here. Firstly, liposomes containing N-glutaryl phosphatidylethanolamine (NGPE) were prepared, followed by the encapsulation of doxorubicin by proton gradient. After the encapsulation of doxorubicin, DH2 or SH1 antibodies were conjugated to NGPE in the liposomes (direct coupling). Secondly, liposomes were prepared with NGPE/mAb conjugates by the detergent dialysis method (conjugate insertion), and then doxorubicin was encapsulated by proton gradient. The immunoliposomes prepared by both methods were able to specifically bind to the surface of the tumor cells - B16BL6 mouse melanoma cells and HRT-18 human colonic adenocarcinoma cells. The efficiencies of doxorubicin-entrapping into liposomes prepared by direct coupling and conjugate insertion was about 98% and 25%, respectively. These types of liposomal formulation are sensitive to target cells, which can be useful for various clinical applications.
Convenient Preparation of Tumor-specific Immunoliposomes Containing Doxorubicin
Park, Yong-Serk,Cho, Jang-Eun,Son, Byoung-Soo,Nam, Sang-Min The Korea Science and Technology Center 1998 BMB Reports Vol.31 No.1
Two innovative methods to prepare target-sensitive immunoliposomes containing doxorubicin by coupling monoclonal antibodies (mAb DH2, SH1) specific to cancer cell surface antigens (Gm₃, Le) have been developed and are described here, liposomes containing N-glutaryl phosphatidylethanolamine (NGPE) were prepared, followed by the encapsulation of doxorubicin by proton gradient. After the encapsulation of doxorubicin, DH2 or SH1 antibodies were conjugated to NGPE in the liposomes (direct coupling). Secondly, liposomes were prepared with NGPE/mAb conjugates by the detergent dialysis method (conjugate insertion), and then doxorubicin was encapsulate by proton gradient. The immunoliposomes prepared by both methods were able to specifically bind to the surface of the tumor cells- B16B6 mouse melanoma cells and HRT-18 human colonic adenocarcinoma cells. The efficiencies of doxorubicin-entrapping into liposomes prepared by direct coupling and conjugate insertion was about 98% and 25%, respectively. These types of liposomal formulation are sensitive to target cells, which can be useful for various clinical applications.
Hyun-Gyu Park,Yong-Seong Lee,Kil-Yong Kim,Yun-Serk Park,Ki-Hyung Park,Tae-Ho Han,Chong-Min Park,Young Sang Ahn 한국원예학회 2017 원예과학기술지 Vol.35 No.1
The objective of this study was to determine whether inoculation with Bacillus licheniformis MH48 as a plant growth-promoting rhizobacterium (PGPR) could promote nutrient uptake of seedlings of the ornamental plant Camellia japonica in the Saemangeum reclaimed coastal land in Korea. B. licheniformis MH48 inoculation increased total nitrogen and phosphorus content in soils by 2.2 and 20.0 fold, respectively, compared to those without bacterial inoculation. In addition, B. licheniformis MH48 produced auxin, which promoted the formation of lateral roots and root hairs, decreased production of growth-inhibiting ethylene, and alleviated salt stress. Total nitrogen and phosphorus uptake of seedlings subjected to bacterial inoculation was 2.3 and 3.6 fold higher, respectively, than the control. However, B. licheniformis MH48 inoculation had no significant effect on the growth of seedlings. Our results suggest that inoculation with B. licheniformis MH48 can be used as a PGPR bio - enhancer to stimulate fine root development, promote nutrient uptake and alleviate salt stress in ornamental plant seedlings grown in the high-salinity conditions of reclaimed coastal land.