• Title/Summary/Keyword: radiolabelling

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Immunoreactivity of Radiolabelled Monoclonal Antibody and Sensitivity of Immunoradiometric Assay: Effect of Labelling Method and Specific Activity (동위원소 표지 단세포군항체의 면역반응성과 방사면역계수법의 예민도 : 표지방법 및 비방사능이 미치는 영향)

  • Ryu, Jin-Sook;Moon, Dae-Hyuk;Cheon, Jun-Hong;Lee, Myung-Hae;Chung, Hong-Keun
    • The Korean Journal of Nuclear Medicine
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    • v.27 no.2
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    • pp.261-269
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    • 1993
  • When monoclonal antibodies are used in radioimmunoassay or immunoscintigraphic studies, post-labelling immunoreativity is a critical parameter. $^{125}I$ was incorporated to CEA-79 (anti CEA monoclonal antibody developed in Korea) by chloramine T and iodogen method with variable specific activities from $0.1{\mu}Ci/{\mu}g$ to $100{\mu}Ci/{\mu}g$. We used a new method to evaluate the immunoreactivites of modified antibody relative to the unlabelled native antibody from competitive binding assay. The effect of immunoreactivity and specific activity to the sensitivity of radioimmunometric assay was also evaluated. As a result, chloramine T method was better than iodogen method in radioiodination of CEA-79, because the immunoreactivity of antibody was relatively well reserved and more stable. New competitive binding assay was simple and effective to evaluate the change of immunoreactivity in radiolabelling. Antibody with high immunoreactivity and high specific activity improved the sensitivity of radioimmunometric assay, whereas antibody with high specific activity but low immunoreactivity didn't. The immunoreactivity and specific activity should be optimized according to the clinical un, and competitive binding method is useful in selection of optimal radiolabelling assay.

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A study on Radiolabelling Method in Radioimmunoassy -Part 2. Determination of Serum Thyrotropin using new Radiolabelling Method- (방사면역측정(放射免疫測定)에서 방사성동위원소(放射性同位元素) 표식법(標識法) 개발에 관한 연구(硏究) -제2편(第2編) 새로운 표식법(漂識法)을 이용한 혈청(血淸) 갑상선자극(甲狀腺刺戟) 홀몬 측정(測定)-)

  • Cho, Bo-Yeon;Suh, Jung-Hun;Park, Yong-Ja;Kim, Byung-Kook;Koh, Chang-Soon
    • The Korean Journal of Nuclear Medicine
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    • v.15 no.2
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    • pp.35-41
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    • 1981
  • The radioimmunoassay of human thyrtropin was performed in various thyroid diseases, using the antin-h-TSH antibody (Daiichi, Japan) and purified h-TSH(Biodata, Italy). $^{125}I$ labelling of h-TSH was performed using a small amount $(1.8{\mu}g)$ of chloramin-T as an oxidant at room temperature. This new method facilitated uniform labelling and reduced to damage of h-TSH by chloramin-T, and satisfactory radioimmunoasasy results were obtained with $^{125}I-TSH$ prepared by the new method until at least 7 weeks after preparation, The assay sensitivity was $1.0{\mu}u/ml$. The coefficient of variances in within assay and interrun assay were all less than 10% among the range of $1.25{\mu}u/ml$ and $160{\mu}u/ml$. The serum TSH values by this new method were colsely related to those by commercial kit (r= 0.996).

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A Study on Radiolabelling Method in Radioimmunoassay -Part 1, A New Method for the $^{125}I-Labelling$ of Human Thyrotropin under Mild Conditions- (방사면역측정(放射免疫測定)에서 방사성(放射性) 동위원소(同位元素) 표식법(標識法) 개발에 관한 연구(硏究) -제1편(第1編). Human Thyrotropin의 온화한 조건하 $^{125}I$-표지를 위한 새로운 방법-)

  • Suh, Jung-Hun;Cho, Bo-Yeon;Park, Yong-Ja;Kim, Byong-Kook;Koh, Chang-Soon
    • The Korean Journal of Nuclear Medicine
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    • v.15 no.2
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    • pp.27-33
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    • 1981
  • Human thyrotropin(h-TSH)의 $^{125}I$-표지를 실온서 소량의 Chloramine T를 산화제로 사용하여 수행하였다. 이 방법은 다량의 Chloramine T를 사용하는 종래의 방법과 비교하여 균일한 방사능 표지를 용이하게 하고 강력한 산화제인 Chloramine T에 의한 h-TSH의 파손을 줄여 주었다. 새로운 방법에 의해 합성된 $h-TSH^*$는 항체에 대한 친화도와 유효기간에 있어서 Daiichi 회사의 상품에 비교하여 손색이 없었다.

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Studies on the Biological Behaviors of Taxol Derivatives (Taxol 유도체들의 생물학적 거동에 관한 연구)

  • Awh, Ok-Doo;Yoo, Dae-Wung;Im, Sang-Moo
    • The Korean Journal of Nuclear Medicine
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    • v.31 no.4
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    • pp.440-451
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    • 1997
  • This study was designed to prospect the $^{111}In$-labelled paclitaxel as tumor imaging agent. In order to provide a taxol molecule with a functional group which is able to chelate In-111, 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 $^{111}InCl_3$ or indirectly with $^{111}In$-citrate(ligand-exchange method). The ligand-exchange method was not suitable because some precipitates appeared during the reaction. On the contrary, by direct radiolabelling method, we were able to obtain taxol-DTPA-$^{111}In$ in 100% radiochemical yield. However, 2'-hemisuccinyltaxol was not labelled by both methods. Yield and radiochemical purity of the radiolabelled com-pound were determined by HPLC, paper chromatography and instant thin layer chromatography. Taxol-DTPA-$^{111}In$ was characterized to be hydrophilic by lipophilicity test, and nearly non-adhesive to HT29, B16, P388, and CT26 by cell binding affinity test. Binding affinity of the taxol-DTPA-$^{111}In$ complex to serum proteins was also examined by protein precipitation with 30% trichloroacetic acid. The results showed that 30% of the taxol-DTPA-$^{111}In$ complex binds with serum proteins.

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Effect of Thermalization and Ultrafiltration Membrane on the Increase of Cottage Cheese Yield Using Radiolabelled Protein (방사성 표지단백질을 이용한 우유의 열처리 및 한외거르기가 코티지 치즈의 생산성 증대에 미치는 영향)

  • Noh, Bong-Soo;Park, In-Seon
    • Korean Journal of Food Science and Technology
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    • v.22 no.7
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    • pp.774-779
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    • 1990
  • $[^{14}C]$-radiolabelled ${\beta}-lactoglobulin$ was used for the studies on the effect of thermalization and ultrafiltration for the increase of cheese yield. 4.33% of ${\beta}-lactoglobulin$ was incorporated through thermalization. $3.20{\sim}3.65%$ of ${\beta}-lactoglobulin$ was more incorporated with cheese curd in the thermalization and ultrafiltration than without ultrafiltration process. Comparing with protein increase, other whey proteins might be incorporated with casein micelles. Loss of $[^{14}]C-{\beta}-lactoglobulin$ through processing and adsorption to membrane during ultrafiltration was only 1.03%.

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Radiochemical separation of 89Zr: a promising radiolabel for immuno-PET

  • Vyas, Chirag K.;Park, Jeong Hoon;Yang, Seung Dae
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.2 no.1
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    • pp.43-50
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    • 2016
  • $^{89}Zr$ with the favorable nuclear decay kinetics and chemical properties is an appealing radiometal for its application in immuno-PET using radiolabeled monoclonal antibodies. Rising demand of ultrahigh purity and high-specific activity $^{89}Zr$ has propelled the radiochemist worldwide to develop an overall efficacious method for its promising separation from the target matrix $^{89}Y$. The requirement of elevated radiochemical purity (${\geq}$ 99.99%) has accelerated the efforts since last two decades to achieve higher decontamination and separation factors of carrier free $^{89}Zr$ over $^{89}Y$ using several suitable separation techniques. However, each of the technique has its own pros and cons which prior to its actual medical application needs to be optimized and thoroughly scrutinized to avoid further complications during radiolabelling of the pharmaceuticals. In this short review article we will specifically consider as well focus on the historical development and the recent advances on the radiochemical separation of $^{89}Zr$ from $^{89}Y$ which will be helpful for the separation scientist involved in this area to understand the existing available means and plan the strategy to investigate and develop the novel techniques to overcome the problems involved in the present methods.

Expression of Human Papillomavirus Type 16, Prototype and Natural Variant E7 Proteins using Baculovirus Expression System

  • Han, Hee-Sung;Kee, Sun-Ho;Hwang, Soon-Bong;Kim, Hyung-Jun;Cho, Kyung-A;Kim, Yoon-Won;Cho, Min-Kee;Chang, Woo-Hyun
    • The Journal of Korean Society of Virology
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    • v.28 no.1
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    • pp.53-62
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    • 1998
  • Human papillomavirus (HPV) 16, E7 proteins derived from the prototype (Bac73) and natural variant (Bac101) E7 open reading frame were produced in Sf9 insect cells. The variant E7 gene occurred naturally by substitution mutation at the position of 88 nucleotide, resulting serine instead of asparagine. Using E7 specific monoclonal antibody (VD6), both E7 proteins were identified in recombinant baculovirus infected SF9 cells. Radiolabelling and immunoprecipitation analysis revealed that both E7 proteins were phosphoproteins. Immunostaining result showed that E7 proteins were mainly localized in the cytoplasm. Nuclear form of E7 proteins was also detected after a sequential fractionation procedure for removing chromatin structure. Considering that the VD6 recognition site in E7 protein is located within 10 amino acid at the N-terminus, this region appears to be blocked by the nuclear component. Western blot analysis revealed that nuclear form was more abundant than cytoplasmic E7 proteins. Time course immunostaining showed that the primary location of E7 protein was the nucleus and exported to the cytoplasm as proteins were accumulated. These events occurred similarly in both Bac73 and Bac101 infected Sf9 cells, suggesting that these two proteins may have similar biological functions.

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Synthesis and Evaluation of a Ligand Targeting the Somatostatin Receptor for Drug Delivery to Tumor Cell (암세포 내로의 약물 전달 증진 목적의 신규 소마토스타틴 수용체 타겟리간드 합성 및 평가)

  • Choi, SunJu;Hong, YoungDon;Lee, SoYoung;Jung, SungHee
    • Journal of Radiation Industry
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    • v.9 no.4
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    • pp.193-198
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    • 2015
  • Most of targeted therapies block the action of certain enzymes, proteins, or other molecules involved in the growth and spread of cancer cells to produce its cytotoxic effect. Either small molecule drugs or monoclonal antibodies are mostly used in targeted therapies. Unfortunately, targeted therapy has a certain degree of unwanted side effect like other cytotoxicity inducing chemotherapies. To overcome and to reduce unwanted side effects during a cancer therapy, recently radiopeptide therapies has got the worlds' attraction for the tumor targeting modalities due to its beneficial effect on less side effect compared to cytotoxic chemotherapies. Among radiopeptide therapies, $^{177}Lu$-DOTATATE is a major modality as an effective one invented so far in treating neuroendocrine tumor (NET) and it has been in clinical trials at least one decade. Although it does have rather effective therapeutic effect on NET, it has less effective in rather large solid tumor. There are many ways to improve or increase therapeutic effect of radiopeptide are a finding the potent small molecules to target the tumor site selectively, or a labeling with radioisotope of emitting high energy, or an improving its biological half-life by introducing different moieties to increase lipophilicity. Present study was focus to increase a biological half-life of radio somatostatin which will target the somatostatin receptor by altering the bifunctional chelator (BFCA) by introducing lipophilic moiety to the somatostatin, which would make the labeled peptide stay longer in the tumor site and thus it can intensify the therapeutic effect on tumor cell itself and around tissues.

Synthesis and biological evaluation of diagnostic reagent for prostate cancer using copper-64 radioisotope

  • Ahn, Heesu;Kim, Mi Hyun;Han, Sang Jin;Woo, Sang Keun;Kim, Jung Young;Lee, Kyu Chul;Lim, Il Han;Lee, Yong Jin
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.4 no.2
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    • pp.65-72
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    • 2018
  • Prostate specific membrane antigen (PSMA) is a cell surface membrane protein, which is overexpressed in most prostate cancer. Recently, PET imaging with $[^{68}Ga]$PSMA-HBED-CC has been widely used for the diagnosis of recurrent prostate cancer and the studies on the diagnostic potential of $^{64}Cu$-labeled PSMA ligands reported actively. In this study, we monitored with biological evaluation in vivo and PET imaging of $^{64}Cu$-labeled PSMA ligand ($[^{64}Cu]$PSMA-617). The radiolabelling efficiency and stability of $[^{64}Cu]$PSMA-617 were confirmed by radio-thin layer chromatography. The radiolabeling efficiency of $[^{64}Cu]$PSMA-617 showed over 95%, and stabilities of intact remained over 98% in both human and mouse serum for 48 h. In normal male mice, in vivo uptake of $[^{64}Cu]$PSMA-617 in several organs was measured at 2, 4, 6, 24, 48 h after injection. Rapid blood clearance was observed for $[^{64}Cu]$PSMA-617. The high uptake was observed in the lung, liver, intestines and kidneys at 2 h postinjection, but was low in the other organs (1-2 %ID/g) at 4 h. The dynamic PET/CT images of 22RV1 tumor-bearing nude mice were acquired during 60 min and additionally acquired 24 h and 48 h after injection. In dynamic PET images, $[^{64}Cu]$PSMA-617 uptake ratio in tumors versus muscle was increased as time elaplsed until 60 minutes and remained in tumors at 48 h. In these results, the PET/CT imaging using $[^{64}Cu]$PSMA-617 in prostate cancer is expected to be useful for the diagnosis and treatment of prostate cancer patients.