• The Korean Journal of Nuclear Medicine
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• v.28 no.1
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• pp.106-111
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• 1994

2-iminothiolane is known to bind $NH_2$ group of lysine in the protein and deliver SH group, which can be used to label protein with $^{99m}Tc$. In this study, we looked for the best reaction condition in which 2-iminothiolane is conjugated to human polyclonal IgG and labeling condition with $^{99m}Tc$-glucoheptonate. Labeling yield was measured with TSK G4000SW column and HPLC or precipitation with 10% TCA (trichloroacetic acid) and 1% HSA. In vivo distribution was investigated with Staphylococcal abscess bearing rats. With decreasing glucoheptonate, the labeling yield decreased. Without 2-iminothiolane, $^{99m}Tc$-glucoheptonate was bound to IgG, which seemed to be direct labeling. With increasing 2-iminothiolane upto 20 times higher than IgG, the labeling yield increased, and plateau was seen with higher molar excess of 2-iminothiolane. Polymer formation was not observed. The pH for the conjugation of 2-iminothiolane and IgG was best around 6.4. $^{99m}Tc$-2-iminothiolane-IgG showed faster blood clearance, higher renal activity and lower hepatic and splenic activity than $^{99m}Tc$-DTPA-IgG. The biodistribution of $^{99m}Tc$-2-iminothiolane-IgG with higher molar excess of 2-iminothiolane was not different from that with lower molar excess. Labeling antibodies with $^{99m}Tc$ using 2-iminothiolane can afford a possible route to simple labeling and wide clinical use of the immunoscintigraphy.

• Nuclear Engineering and Technology
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• v.34 no.2
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• pp.146-153
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• 2002

To radiolabel bioactive molecules, we synthesized $^{99m}$Tc-tricarbonyl precursor, [$^{99m}$Tc(CO)$_3$($H_2O$)$_3$]$^{+}$ with a low oxidation state ( I ). The [$^{99m}$Tc(CO)$_3$($H_2O$)$_3$]$^{+}$ was prepared by low pressure carbonylation (1 atm of CO) of [$^{99m}$Tc $O_4$)]$^{[-10]}$ in the presence of NaB $H_4$ resulting in higher than 98% of labeling yield and stability up to 8 hrs. We evaluated the characteristics of $^{99m}$Tc- tricarbonyl labeled bioactive molecules by carrying out in vitro and in vitro study. Prepared [$^{99m}$Tc(CO)$_3$($H_2O$)$_3$]$^{+}$ was then reacted with some ligands of significance in modem diagnostic nuclear medicine and some amino acids. Labeling yields were checked by HPLC and found to be usually high, excluding $^{99m}$Tc-tricarbonyl-MDP, -EDTMP and -mIBG. And the biodistribution properties of $^{99m}$Tc-tricarbonyl complexes applied in rabbit showed different appearance comparing with that of the $^{99m}$Tc-labeling by conventional means. From these results, we conclude that [$^{99m}$Tc(CO)$_3$($H_2O$)$_3$]$^{+}$ is a potential precursor for development of radiopharmaceuticals, especially for labeling of biomolecules.

• The Korean Journal of Nuclear Medicine
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• v.27 no.2
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• pp.270-276
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• 1993

$Technetium-^{99m}$ labeling method using bifunctional chelating agent cyclic DTPA has been evaluated with human polyclonal nonspecific IgG. IgG was conjugated with cyclic DTPA with various molar ratio. Reduction of $^{99m}Tc$ was done with $Na_2S_2O_4$ with various molar excess. Labeling efficiency and identification of polymer was confirmed with HPLC using TSK4000 SW column. Polymer was purified with 100 cm Sepharose 6LB column. Cultured $1{\times}10^9$ Staphylococcus aureus were injected into rat thigh 24 hours later labeled IgG was injected, and in vivo distribution was observed 4 and 24 hours thereafter. Reduction of $^{99m}Tc$ was optimal with the 10000-50000 times molar excess of $Na_2S_2O_4$. Polymer formation increased with increasing mloar excess of cyclic DTPA to IgG. Three step labeling-labeling DTPA conjugated IgG after reduction of $^{99m}Tc$-made more polymer than two two step labeling-simultaneous mixing DTPA conjugated IgG, $^{99m}Tc$ and $Na_2S_2O_4$. $^{99m}Tc$ blood clearance and lower uptake in the abscess and other organs. IgG conjugated with 200 times molar excess of cyclic DTPA showed slower blood clearance with 200 times molar excess of cyclic DTPA showed slower blood clearance than that of 200 times molar excess of cyclic DTPA showed slower blood clearance than that of 20 times molar excess. In the $^{99m}Tc$ labeling of IgG with cyclic DTPA for the immunoscintigraphy, obtimal labeling condition should be chosen, and effect of the $^{99m}Tc$ labeled IgG polymer should be considered.

• The Korean Journal of Nuclear Medicine Technology
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• v.16 no.2
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• pp.131-134
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• 2012

Purpose : The labeling efficiency of radiopharmaceuticals in nuclear medicine is important in terms of accuracy and reliability of the examination. Usually $^{99m}Tc$-HMPAO used for brain SPECT scan is chemically unstable since lots of impurities are existing. Therefore, occurrence of loss of labeling efficiency is easy to appear. In this paper, labeling and use of $^{99m}Tc$-HMPAO should be helpful through experiments on factors affecting the labeling efficiency of $^{99m}Tc$-HMPAO. Materials and Methods : Domestic HMPAO vials (Dong-A) used for brain SPECT scan were tested. Domestic Samyeong Generator 55.5 GBq (1,500 mCi), TLC measurement sets (ITLC-SG, butanone, saline, TLC chamber) and radio-TLC scanner (Advantest, Bioscan) were used. In the first experiment, after eluting generator at 1, 8, 16, 24, 28 hours apart, each eluted $^{99m}Tc$-pertechnetate were labeled with HMPAO and the labeling efficiency was measured. In the second experiment, after eluting $^{99m}Tc$-pertechnetate from a generator, $^{99m}Tc$-pertechnetate was drawn at 0, 1, 3, 6 hours. And each drawn $^{99m}Tc$-pertechnetate were labeled with HMPAO for measuring labeling efficiency. In the third experiment, labeling efficiency was measured at 0, 0.5, 3, 5, 7 hours after labeling $^{99m}Tc$-HMPAO. Results : In the first experiment, measured values were appeared 95.05, 94.64, 94.94, 95.64, 96.76% in passing order of time. In the second experiment, measured values were appeared 94.38, 94.23, 93.26, 91.03% in passing order of time. In the third experiment, measured values were appeared 95.76, 94.17, 88.19, 83.6, 76.86% in passing order of time. Conclusion : In the first experiment of this paper, labeling efficiency of $^{99m}Tc$-HMPAO labeled with $^{99m}Tc$-pertechnetate eluted after 24 hours from first elution. Additional experiments will be needed to discuss for usability. In the second experiment, the labeling efficiency was slightly decreased in chronological order, but it was measured higher than 90%. Also, additional experiments will be needed to discuss for usability. In the third experiment, the labeling efficiency was decreased considerably. Especially, within 3 hours after the labeling is recommended to use $^{99m}Tc$-HMPAO

• The Korean Journal of Nuclear Medicine
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• v.30 no.3
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• pp.361-366
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• 1996

This study was to determine the effect of $Al^{3+}$ in $^{99m}Tc$ eluate from $^{99}Mo-^{99m}Tc$ generator on labeling efficiency and biodistribution of $^{99m}Tc$-MDP. The chromatographic analysis of $^{99m}Tc$-MDP preparations containing $Al^{3+}(0-62.5{\mu}g/ml)$ showed decreased labeling efficiency $^{99m}Tc$ pertechnetate and hydrolyzed reduced $^{99m}Tc$ fraction increased with increasing concentrations of aluminum. However, the chromatography system could not discern between hydrolyzed reduced $^{99m}Tc$ and $^{99m}Tc$ labeled colloid. $^{99m}Tc$-MDP preparations containing aluminum were relatively stable. Chromatographic analysis also confirmed that no significant differences were observed in the radiochemical purity of the filtered and the unfiltered $^{99m}Tc$-MDP preparations containing aluminum by $0.22{\mu}m$ syringe filter. In biodistribution data of ICR-mice, blood and heart uptake were increasing with increasing concentrations of aluminum, because of decreasing labeling efficiency of $^{99m}Tc$-MDP and increasing of $^{99m}Tc$ pertechnetate. However, liver and bone uptake were not significantly increased. In rat images no difference were observed at $5{\mu}g/ml\;Al^{3+}$ compare with at $0{\mu}g/ml\;Al^{3+}$, but at $10{\mu}g/ml\;Al^{3+}$ lumbar uptake was increased. As a practical conclusion, a concentration below $10{\mu}g/ml\;Al^{3+}$($10{\mu}g/ml\;Al^{3+}$ is the maximum allowed in pertechnetate eluate from $^{99}Mo-^{99m}Tc$ generator by USP.) in $^{99m}Tc$-MDP radiopharmaceutical result in low labeling efficiency. Radiochemical purity 90% of $^{99m}Tc$-MDP is the minimum allowed by USP. Therefore, when soft tissue uptake is observed in $^{99m}Tc$-MDP bone scan and labeling efficiency is above 90%, we can expect that $Al^{3+}$ in pertechnetated eluate is not the cause of soft tissue uptake.

• Archives of Pharmacal Research
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• v.27 no.9
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• pp.961-967
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• 2004

The development of an antibody labeling method with $^{99m}$Tc is important for cancer imaging. Most bifunctional chelate methods for $^{99m}$Tc labeling of antibody incorporate a $^{99m}$Tc chelator through a linkage to lysine residue. In the present study, a novel site-specific $^{99m}$Tc labeling method at carbohydrate side chain in the Fc region of 2 antibodies (T101 and rabbit anti-human serum albumin antibody (RPAb)) using dihydrazinophthalazine (DHZ) which has 2 hydrazino groups was developed. The antibodies were oxidized with sodium periodate to pro-duce aldehyde on the Fc region. Then, one hydrazine group of DHZ was conjugated with an aldehyde group of antibody through the formation of a hydrazone. The other hydrazine group was used for labeling with $^{99m}$Tc. The number of conjugated DHZ was 1.7 per antibody. $^{99m}$Tc labeling efficiency was 46-85％ for T101 and 67∼87％ for RPAb. Indirect labeling with DHZ conjugated antibodies showed higher stability than direct labeling with reduced antibodies. High immunoreactivities were conserved for both indirectly and directly labeled antibodies. A biodistribution study found high blood activity related to directly labeled T1 01 at early time point as well as low liver activity due to indirectly labeled T101 at later time point. However, these findings do not affect practical use. No significantly different biodistribution was observed in the other organs. The research concluded that DHZ can be used as a site-specific bifunctional chelating agent for labeling antibody with $^{99m}$Tc. Moreover, $^{99m}$Tc labeled antibody via DHZ was found to have excellent chemical and biological properties for nuclear medicine imaging.edicine imaging.

• Journal of Radiopharmaceuticals and Molecular Probes
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• v.3 no.1
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• pp.38-43
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• 2017

Tc-99m labeled sestamibi ($^{99m}Tc$-MIBI) is one of most widely used radiopharmaceuticals for myocardial SPECT imaging. Radiolabeling of $^{99m}Tc$-MIBI is recommended by heating in $100^{\circ}C$ water bath for 15 min. However, the water bath might be a source of contamination. Thus, if radiolabeling of $^{99m}Tc$-sestamibi can be performed at room temperature, then it would be more convenient to use in clinical application. In this study, we performed the radiolabeling of $^{99m}Tc$-MIBI in different temperature conditions or using different instruments to find out the efficient labeling condition. We studied the $^{99m}Tc$-MIBI labeling at room temperature or $100^{\circ}C$ heating block, and checked the labelling yields every 1 min for 10 min using radio-TLC with 2 different eluents-saline and acetone. From the experiment, we confirmed that the $^{99m}Tc$-MIBI can be labeled over 90% yield but not completed at room temperature. However, the $^{99m}Tc$-MIBI labeling was completed when it was performed in the $100^{\circ}C$ heating block. Finally, we proved that heating is essential for complete $^{99m}Tc$-MIBI labelling, furthermore using heating block is also possible instead of water bath.

• The Korean Journal of Nuclear Medicine Technology
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• v.14 no.1
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• pp.90-93
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• 2010

Purpose: In our nuclear medicine department, we suggested AMC RBC labeling method improved by modifying a part of existing modified in-vitro method to raise the efficiency of $^{99m}Tc$-RBC labeling. However, it needs to be more additional time and efforts than existing modified in-vitro method because the AMC RBC labeling method has to carry out the centrifugal separation process for 3~5 minutes. Therefore, in this study, we conducted researches to aim to maintain stable labeling effects and supplement a problem about additional time by reducing rotating time when labeling $^{99m}Tc$-RBC. Materials and Methods: This research has been conducted the object of 30 patients who examined study using $^{99m}Tc$-RBC and agreed to this research at our hospital from May 2009 to September 2009. We made 4 blood samples which consisted of ACD 1 cc along with 5 cc blood from each patient and used the AMC RBC labeling method. At this moment, each labeling efficiency was calculated by different rotating time 5 min, 10 min, 15 min, and 20 min and then we compared differences. Results: As a result, When comparing the $^{99m}Tc$-RBC labeling method efficiency by using the AMC RBC labeling method which differents from rotating time, each labeling efficiency were $92.3{\pm}5.0%$ in 5 min, $95.9{\pm}5.0%$ in 10 min, $97.4{\pm}4.9%$ in 15 min and $97.7{\pm}4.8%$ in 20 min. We analyzed differences of the labeling efficiency from change of rotating time by using an one-way ANOVA and verified that in Duncan method. There was relatively efficiency low in 5min rotating time and no statistically significant change in over. Conclusions: When comparing a existing method, the AMC RBC labeling method which goes through the centrifugal separation process again offers more favorable condition to combine RBC with $^{99m}TcO4^-$ by eliminating an plasma ingredient. When using the modified in-vitro method, we have almost 20 min to rotate to acquire stable labeling efficiency. But, when using the AMC RBC labeling method, we acquire labeling efficiency well what we want within only 10 min to rotate. Decrease of rotating time can complement the AMC RBC labeling method which goes through the centrifugal separation process again and also provide more rapid study such as G-I bleeding study due to fast labeling.

• Journal of Microbiology and Biotechnology
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• v.18 no.9
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• pp.1599-1605
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• 2008

Biodistribution and lymphoscintigraphy of cyclosporine A (CyA) and technetium-99m ($^{99m}Tc$) were studied using ${^99m}Tc$-labeled dextran acetate (DxA) including CyA. DxA particles were prepared from dextran with acetic anhydride, and CyA was loaded into them. Lymphatic delivery of ${^99m}Tc$-labeled DxA particles containing CyA was evaluated after subcutaneous injection into the foot pad of rats and compared with those of ${^99m}Tc$-labeled human serum albumin (HSA). The labeling efficiency of CyA-loaded ${^99m}Tc$-DxA particles was about 95% at 30 min. The labeling efficiency maintained stably above 80% for 12 h. The percent injected dose (%ID) of CyA-loaded ${^99m}Tc$-DxA was similar to that of ${^99m}Tc$-HSA at the inguinal lymph node after 40 min. The CyA-loaded ${^99m}Tc$-DxA could be as well distributed as ${^99m}Tc$-HSA through the lymph node. The DxA particles could steadily distribute the CyA as well as the ${^99m}Tc$ radiolabeling through the lymph node.

• The Korean Journal of Nuclear Medicine
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• v.29 no.1
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• pp.110-117
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• 1995

Tc-99m Labeled hexamethylenepropyleneamineoxime ([$^{99m}Tc$]-HMPAO) is a famous amino-oxime compound and is widely used to construct SPECT images of cerebral blood flow. To investigate the relationship between chemical structure and radiolabeling in these kind of diamine-oxime compounds, we synthesized seven compounds by Schiff's base formation and successive reduction with sodium borohydride. They were (RR/SS )-4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione bisoxime (2), (RR/SS/meso)-4,8-diaza-3,9-dimethy-lundecane-2,10-dione bisoxime (4), (RR/SS/meso)-4,8-diaza-3,10-dimethyldodecane-2,11-dione bisoxime (5), (RR/SS/meso)-4,7-diaza-3,6,6,8-tetramethyldecane-2,9-dione bisoxime (8), (RR/SS/meso)-4,7-diaza-5,6-cyclohexyl-3,8-dimethyldecane-2,9-dione bisoxime (10), (RR/SS/meso)-3,4-bis(1-aza-2-methyl-3-oxime-1-butyl)-benzoic acid (12), and (RR/SS/ meso)-2,3-bis(1-aza-2-methyl-3-oxime-1-butyl) benzophenone (14). Chemical structures of all the synthesized compounds were identified by taking $^1H$ spectrum. Among them, 2 and 4 are propyleneamine oxime (PnAO), 6 is butyleneamine oxime (BnAO) and 8, 10, 12 and 14 are ethyleneamine oxime (EnAO). Each compound (0.5 mg) was incubated with stannous chloride (0.5 g - 8 g), carbonate-bicarbonate buffer (final concentration = 0.1 M, pH 7 - pH 10) and Tc-99m-pertechenate (1 ml). Tc-99m labeling of these compounds were checked by ITLC (acetone), ITLC (normal saline), reverse phase TLC (50 % acetonitrile) and ITLC (ethyl acetate). According to the results, EnAO's were not labeled by Tc-99m in any of above condition. About 11 % of maximum labeling efficiency was obtained with BnAO. However, 4 (PnAO) was labeled with Tc-99m to 85 % which is similar to the labeling efficiency of 2 (HMPAO). Hydrophilic impurity (9 % ) was the most significant problem with the labeling of 4, however, pertechnetate (3 % ) and colloid (3 %) were minor problem. In conclusion, we synthesized seven diamine blsoxlme compounds. Among them, four EnAO compounds were not labeled by Tc-99m. A BnAO was labeled poorly and two PnAO's were labeled well. These labeling can be explained by tertiary structure of their Tc-99m chelate.