Sutari Sutari
Center for Radioisotopes and Radiopharmaceuticals, National Nuclear Energy Agency Puspiptek Area, Serpong, 15314, Indonesia

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Synthesis of 125I Labeled Estradiol-17β-Hemisuccinate and Its Binding Study to Estrogen Receptors Using Scintillation Proximity Assay Method Susilo, Y.; Mondrida, G.; Setiyowati, S.; Sutari, Sutari; Triningsih, Triningsih; Lestari, W.; Widayati, P.; Ardiyatno, C.N.; Ariyanto, A.; Darwati, S.; Kardono, L.B.S.; Yanuar, A.
Atom Indonesia Vol 38, No 3 (2012): December 2012
Publisher : PPIKSN-BATAN

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1276.418 KB) | DOI: 10.17146/aij.2012.175

Abstract

Research was carried out to obtain a selective ligand which strongly bind to estrogen receptors through determination of binding affinity of estradiol-17β-hemisuccinate. Selectivity of these compounds for estrogen receptor was studied using Scintillation Proximity Assay (SPA) method. Primary reagents required in the SPA method including radioligand and receptor, the former was obtained by labeling of estradiol-17β-hemisuccinate with 125I, while MCF7 was used as the receptor. The labeling process was performed by indirect method via two-stage reaction. In this procedure, first step was activation of estradiol-17β-hemisuccinate using isobutylchloroformate and tributylamine as a catalist, while labeling of histamine with 125I was carried out using chloramin-T method to produce 125I-histamine. The second stage was conjugation of activated estradiol-17β-hemisuccinate with 125I-histamine. The product of estradiol-17β-hemisuccinate labeled 125I was extracted using toluene. Furtherly, the organic layer was purified by TLC system. Characterization of estradiol-17β-hemisuccinate labeled 125I from this solvent extraction was carried out by determining its radiochemical purity and the result was obtained using paper electrophoresis and TLC were 79.8% and 84.4% respectively. Radiochemical purity could be increased when purification step was repeated using TLC system, the result showed up to 97.8%. Determination of binding affinity by the SPA method was carried out using MCF7 cell lines which express estrogen receptors showed the value of Kd at 7.192 x 10-3 nM and maximum binding at 336.1 nM. This low value of Kd indicated that binding affinity of estradiol-17β-hemisuccinate was high or strongly binds to estrogen receptor.Received: 04 December 2012; Revised: 19 December 2012; Accepted: 21 December 2012
PRODUCTION OF IMMUNORADIOMETRICASSA Y (IRMA) CA 15.3 KIT COMPONENT FOR DETECTION OF BREAST CANCER Widayati, Puji; Ariyanto, Agus; Sutari, Sutari; Mondrida, Gina; Darwati, Siti
Jurnal Radioisotop dan Radiofarmaka Vol 11 (2008): Jurnal PRR 2008
Publisher : Jurnal Radioisotop dan Radiofarmaka

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Abstract

PRODUCTION OF IMMUNORADIOMETRICASSA Y (IRMA) CA 15.3 KITCOMPONENT FOR DETECTION OF BREAST CANCER. YKI has ranked that breast cancer as a second deseas in causing death of Indonesian women (, 2007). This phenomenon has been caused by the low level of public awareness in early detection of cancers. As a consequences this kind of cancer has generally been diagnosed in advanced stadium which is difficult to be treated  In spite of this, the disease actually can be detected early by measuring level of CA 15.3, a tumor marker for breast cancer. One of such in vitro method is immunoradiometricassay (IRMA) for CA 15.3. The CA 15.3 itself is a glycoprotein of heterogen compound capable of reacting with monoclonal antibody CA 15.3. Production of the IRMA CA 15.3 kit has been performed in the Center for Radioisotope and Radiopharmac.eutical, National Nuclear Energy Agency of Indonesia. Optimization of the kit component has been carried out using several parameter including type ofmonoclonal antibody for tracer and buffer coating type. The results showed that monoclonal anti CA M37901M type is better than M37552M type for tracer production. The M3790lM gave yield about 79.51%, with specific activity 29.12 ~Ci/~g, radiochemical purity 94.21% and %B/T about11.94%. Several buffers have been evaluated and 0.05 M pH 9.6 carbonate bicarbonate buffer showed the highest specific binding when it was used as coating buffer. Preparation of IRMA CA 15.3 standard solution gave a linier relation between CA 15.3 concentration and the maximumbinding (%B/T) Y=0.227X+0.5177 and correlation coefficient R 0.9840Keywords: Radioimmunoassay, Immunoradiometricassay, tumor marker, CA-15.3
OPTIMASI RANCANGAN ASSAY KIT IRMA CA-125 Widayati, Puji; Ariyanto, Agus; Abidin, Zaenal; Yunita, F.; Sutari, Sutari
Jurnal Radioisotop dan Radiofarmaka Vol 9 (2006): jurnal PRR 2006
Publisher : Jurnal Radioisotop dan Radiofarmaka

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Abstract

OPTIMASI RANCANGAN ASSAY KIT IRMA CA-125. Carbohydrate Antigen-125 (CA-125) adalah antibodi yang bereaksi spesifik dengan monoklonal CA-125. Antigen CA-125 dapat digunakan sebagai tumor marker dan penentuan kadarnya dapat dilakukan dengan teknik Immunoradiometricassay (IRMA). Pusat Pengembangan Radiosotop dan Radiofarmaka (P2RR-BATAN) telah membuat kit IRMA CA-125 untuk memenuhi kebutuhan dalam negeri. Telah dillakukan optimasi rancangan assay kit IRMA CA-125 buatan PRR tersebut, meliputi penetapan jumlah cacahan perunut, volume perunut, volume standar, waktu inkubasi dan suhu inkubasi yang terbaik sehingga diperoleh nilai %B/T dan % NSB yang optimum dan dapat digunakan sebagai acuan setiap kali assay. Hasil penelitian menunjukkan bahwa jumlah cacahan perunut terbaik adalah ± 100000 cpm, volume perunut terbaik adalah 50µL, volume standar terbaik adalah 50µL, waktu inkubasi terbaik adalah 16 jam dan suhu inkubasi terbaik adalah 25°C (suhu kamar). Penelitian optimasi assay kit IRMA CA-125 menyimpulkan bahwa dengan menggunakan komposisi pereaksi dan kondisi reaksi optimum dihasilkan nilai %B/T = 19,05% dan NSB = 0,53%. Kata kunci: Radioimmunoassay, Immunoradiometricassay, tumor marker, CA-125 ASSAY DESIGN OPTIMIZATION OF CA-125 IRMA KIT. Monoclonal Antibody CA-125 is an antibody against the Carbohydrate Antigen (CA-125). The CA-125 antigen can be used as a tumour marker, and its concentration can be determined by the Immunoradiometricassay (IRMA) method. The center for Radioisotopes and Radiopharmaceuticals (BATAN) has prepared the CA-125 IRMA kit for domestic use. This reportdiscuse the assay design optimization of the a bove mentioned CA-125 IRMA kit, covering determenation of total count oftracer, tracer volume, standart volume, optimum incubation time and temperature, that will be used in daily assay. Investigation showed that the optimum total countof tracer was approx ±100.000 cpm, optimation tracer volume was 50 µL, optimum standard volume was 50 µL, and the optimum incubation time and temperature were 16 hours and 25 °C respectively. It can be concluded that the optimum reagents compositation and reaction conditions can produce %B/T value of 19.05% and NSB value of 0.53%. Key words:Radioimmunoassay, Immunoradiometricassay, tumor marker, CA-125
PEMBUATAN HIT RADIOLVIVIUNOASSAY (RIA) MIKROALBUMINURIA DENGAN METODE COATED TUBE : PERBANDINGAN TABLING POLISTIREN "PLAIN" DAN "DASAR BINTANG" Susilo, V. Yulianti; Sutari, Sutari
Widyariset Vol 12, No 1 (2009): Widyariset
Publisher : LIPI-Press

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Abstract

Microalbuminuria is a physiological condition ofp atient in which albumin is excreted in urine at 20-200 pg/min or 30-300 mg/day. Determination of microalbumin content in the patient urine is very important for early detection ofmicroalbuminuria before nephropathy occurred. Detection ofrnicroalburnin in the urine is commonly performed by radioimmunoassay (R1A) technique. Human serum albumin labeled with 1251 is commonly used as a tracer and bound and unbound phases in the reaction mixture that can be separated easily and quickly with a coated tube method. Production of R1A kit of microalbuminuria  involves preparation of antibody coated into two solid phases of polystyrene tube plain and polystyrene bottom star tube, HSA-251 tracer and standard solution. The polyclonal anti-human serum albumin (anti-HSA)  generated in the rabbit with titer 1 : 15000 was obtained after one year then was coated into polystyrene tubes, the tracer HSA-251 was prepared using an iodogen method. Series of HSA standard were made by dilution of HSA with synthetic urine.  The yield of HSA-121 tracer was about 83,94% with radiochemical purity of 92% and specific activity of 3,3576 pCi/pg. The microalbuminuria RIA kit shows good assay performance of 1, 05% NSB (Non Spesific Binding) and 78,6% B/T The polystyrene bottom star tube shows more higher BIT of bound and could be maintained within 70 days.
OPTIMASI PEMBUATAN COATED TUBE HUMAN SERUM ALBUMIN (HSA) UNTUK KIT RADIOIMMUNOASSAY (RIA) MIKROALBUMINURIA Sutari, Sutari; S., V. Yulianti; Triningsih, Triningsih; Mondrida, Gina; Ariyanto, Agus; Setiyowati, Sri; Widayati, Puji; Lestari, Wening
Jurnal Forum Nuklir JFN Vol 8 No 1 Mei 2014
Publisher : BATAN

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Abstract

OPTIMASI PEMBUATAN COATED TUBE HUMAN SERUM ALBUMIN (HSA) UNTUK KIT RADIOIMMUNOASSAY (RIA) MIKROAMBUMIN. Radioimmunoassay (RIA) adalah suatu metoda analisis berdasarkan pada reaksi imunologi yakni ikatan antigen-antibodi, metoda ini sangat spesifik dan peka digunakan untuk menentukan kadar zat-zat yang ada di dalam cairan tubuh seperti serum, urine dan lainnya sehingga dapat digunakan untuk mengevaluasi suatu penyakit metabolik seperti diabetes melitus. Penelitian dan pengembangan teknologi Kit RIA mikroalbuminuria dengan metode coated tube dilakukan melalui beberapa tahap yakni optimasi pembuatan komponen kit, optimasi assay, validasi assay dan uji klinis. Telah dilakukan penelitian tentang optimasi pembuatan coated tube HSA salah satu komponen kit RIA mikroalbuminuria untuk pemisah fasa padat. Tujuan dari penelitian ini adalah untuk mendapatkan coated tube HSA yang dapat menghasilkan % B/T yang optimum dengan % NSB yang minimum dan memenuhi persyaratan untuk assay. Penelitian dilakukan dengan cara melakukan optimasi larutan dapar sebagai pelarut poliklonal antibodi (Pab)-HSA, optimasi volume coaling (volume Pab-HSA) dan optimasi konsentrasi larutan blocking. Hasilnya menunjukkan bahwa larutan dapar karbonat bikarbonat 0,05 M pH 9,6 memberikan hasil yang optimum sebagai pelarut Pab-HSA pada titer 1:3000, volume Pab-HSA 750 µL dengan 750 µL larutan bovine serum albumin (BSA) 1 % sebagai blocking dan diperoleh % BIT dan % NSB masing-masing 46,49% ± 0,57 dan 0,77% ± 0,04 serta memenuhi persyaratan Kit RIA untuk assay.
Optimization of Thyroglobulin Coated Tube for Thyroglobulin IRMA Kit Sutari, Sutari; Triningsih, Triningsih; Setiyowati, Sri; Susilo, V.Y.; Ariyanto, Agus; Widayati, Puji; Lestari, Wening
JKPK (Jurnal Kimia dan Pendidikan Kimia) Vol 3, No 2 (2018): JKPK (Jurnal Kimia dan Pendidikan Kimia)
Publisher : Program Studi Pendidikan Kimia FKIP Universitas Sebelas Maret

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20961/jkpk.v3i2.22400

Abstract

Immunoradiometric assay (IRMA) is a method of analysis based on immunological reactions of antigens-antibodies binding. This highly specific and sensitive method was used for in vitro diagnosis in small quantity of sample. Center for Radioisotope and Radiopharmaceutical Technology, BATAN has developed Thyroglobulin IRMA Kit using coated tube method that can determine thyroglobulin levels in microgram quantities. Coated tube was made with immobilisation of anti thyroglobulin into polistyrene tube. Development of IRMA kit performed through several steps including: optimization component of kit, optimization assay and kit validation. Optimization of coated tube involved selection and volume of solvent, using blocking and non-blocking agent, and volume of blocking agent. The optimum condition for coated tubes was found to be using 0.1M phosphate buffer pH 7.4 with coating volume of 500 μL, 3% BSA in 500 μL blocking agent 0.1M phosphate buffer pH 7.4, with maximum binding and non-specific binding (NSB) of 60.58 and 1.40%, respectively. The optimized coated tube was found to be stable up to 4 weeks.
Validation of the TSH IRMA Kit for Determination of the TSH Levels in Human Blood Serum Mondrida, Gina; Sutari, Sutari; Triningsih, Triningsih; Setyowati, Sri; Yulianti S, V.; Lestari, Wening; Ariyanto, Agus; Widayati, Puji
JKPK (Jurnal Kimia dan Pendidikan Kimia) Vol 3, No 3 (2018): JKPK( Jurnal Kimia dan Pendidikan Kimia)
Publisher : Program Studi Pendidikan Kimia FKIP Universitas Sebelas Maret

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20961/jkpk.v3i3.22334

Abstract

TSH IRMA kit is a kit used for the determination of TSH (Thyroid Stimulating Hormone) levels in human blood serum. Thyroid hormone is a hormone that our bodies need for growth of the brain, bone and other tissues and regulate the metabolism in the body. TSH normal range for adult is in the range of 0.4-4.5 mIU/L, whereas for baby is about 3.0-18.0 mIU/L. Thyroid would affect the quality of optimal growth of children if disturbed. Therefore, TSH assay in the blood needs to be determined to know whether the function of the thyroid gland works normally or not. Detection of TSH in blood can be performed by Immunoradiometricassay (IRMA) method. IRMA method is one of the immunoassay techniques based on immunological reactions (antigen-antibody binding) using radionuclide 125I as a tracer, that sample in small quantity can be detected.  IRMA method was developed locally by replacing TSH IRMA kit which is costly since imported from commercial companies. Center for Radioisotope and Radiopharmaceutical Technology (PTRR) BATAN has successfully developed the TSH IRMA kit that can be used to determine the levels of TSH in human blood. TSH IRMA kit must be validated to know the limit of detection, sensitivity, accuracy, precision and the assay parameters, such as Non-Specific Binding (NSB) and Maximum Binding (MB). Validation of TSH IRMA kit had been carried out resulting in the limit of detection of 0.115 ng/mL, accuracy with a recovery of 93.6-108.0 %, intra-assay precision (% CV) QC L = 1.9848, QC M = 3.6360 % and QC H = 2.2085 % while the inter-assay precision (% CV) QC L = 11.0055, QC M = 5.6768 %  and  QC H = 5.4181 %.  It was concluded that this TSH IRMA kit showed good performance based on the % NSB and % B/T of 0.68 and 34.64 %, respectively.
UJI BANDING KIT RIA T3 PRODUK PRR-BATAN SISTEM "COATED TUBE" DENGAN PRODUK IZOTOP-HUNGARIA Triningsih, Triningsih; Widayati, Puji; Sutari, Sutari; Setiyowati, Sri
Jurnal Forum Nuklir JFN Vol 8 No 2 November 2014
Publisher : BATAN

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Abstract

UJJ BANDING KIT RIA T3 PRODUK PRR-BATAN SISTEM "COATED TUBE" DENGAN PRODUK IZOTOP-HUNGARlA. Radioimunoassay (RIA) merupakan teknik analisis yang didasarkan pada prinsip immunologi dan menggunakan perunut radioaktif sehingga cuplikan dalam jumlah kecil mudah dideteksi serta spesifik karena didasarkan pada reaksi imunologi yaitu ketika terjadi ikatan antigen antibodi yang spesilik hanya untuk antigen tertentu saja. Teknik ini dapat digunakan untuk penemuan kadar T3 dalam serum yang mempunyai matriks yang komplek dan kadar yang sangat bervariasi. Triiodotironin (T3) adalah salah satu hormon yang diekskresikan oleh kelenjar tiroid. Telah dilakukan penelitian tentang uji banding Kit RIA T3 produk PRR-BATAN sistem "coated tube” dengan produk lzotop-Hungaria sebagai Gold Standard, tujuannya untuk melihat kesesuaian pengukuran kadar T3 dengan cara membandingkan hasil analisis menggunakan kit RIA T3 produk PRR-BATAN sistem “coated tube” dengan produk lzotop Hungaria terhadap 273 sampel yang berasal dari poliklinik PTKMR-BATAN Pasar Jum’at. Hasil uji assay dari kedua kit tersebut diperoleh 185 sampel negatip hipotiroid (true negative), 30 sampel positip hipotiroid (true positive), 17 sampel false positip hipotiroid dan 41 sampcl false negatip hipotiroid. Sedang untuk sampel hipetiroid diperoleh hasil 207 sampel negatip hipertiroid (true negative), 41 sampel positif hipertiroid (true positive), 20 sampel false positip hipertiroid, dan 5 sampel false negatip hipertiroid. Hasil uji banding kit RIA T3 didapatkan nilai diagnostic sensitivity sebesar 42.25% dan diagnostic specivicity sebesar 91.58 % untuk pengukuran sampel hipotiroid, didapatkan diagnostic sensitivity sebesar 89,13% dan diagnostic specivicity sebesar 91,18 % untuk pengukuran sampel hipertiroid sehingga kualitas Kit RIA T3 produk PRR-BATAN sistem "coated tube" belum sama dengan produk dari Izotop Hungaria.Kaw Kunci: Kit, T3, RIA, uji banding
Preparasi Pereaksi Kit Immunoradiometricassa (IRMA)Thyroglobulin (TGB) Untuk Deteksi Kanker Tiroid Widayati, Puji; Ariyanto, Agus; Setiyowati, Sri; Sutari, Sutari; Yulianti, V.
Prosiding Seminar Nasional Teknik Kimia "Kejuangan" 2019: PROSIDING SNTKK 2019
Publisher : Prosiding Seminar Nasional Teknik Kimia "Kejuangan"

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Abstract

In Indonesia thyroid cancer ranks 9th out of 10 malignancies that are often found, in the endocrine system, 1% of all malignancies that exist. Thyroglobulin is a tumor marker for thyroid cancer produced by follicular thyroid cells. Thyroglobulin should not be found in the blood serum of patients after total ablation, but in fact there is still detected thyroglobulin caused by the remaining tumor residue. Measuring levels of TGB was found in the blood can be done by several methods such as by immunoradiometricassay (IRMA) methods or ELISA methods. IRMA method is one of immunoassay techniques using 125I radionuclides  as a tracer,   so the sample in small quantity can be detected. The purpose of this study was obtained TGB reagent kit that includes 125I labeled TGB as a tracer, TGB coated tube and TGB standard of the kit, then it can be optimized assay design, finally TGB reagent kit can be used for early detection of thyroid cancer. Labeling of MAb has been done using 125I with reaction time of 60 seconds.The amount of TGB MAb was 16 μg, 10 μg chloramine T and activity of Na-I125was 1000 μCi. Preparation of TGB coated tube was using phosphate buffer 0,025M pH 7,4 with volume 500 μL, standard TGB 0,1 M phosphate buffer pH 7.4 containing 5% BSA and 0.1% NaN3and resulting of NSB and %B/T were 1,73% and 59,44 % respectifely wich as meet as arequerement of the kit
Rituximab Iodination Procedure for Radioiodinated Rituximab (131I-Rituximab) Preparation Ramli, Martalena; Hidayat, Basuki; Sutari, Sutari; Setyowati, Sri; Susilo, Veronica Yulianti
Majalah Kedokteran Bandung Vol 51, No 2 (2019)
Publisher : Faculty of Medicine, Universitas Padjadjaran

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1864.956 KB) | DOI: 10.15395/mkb.v51n2.1595

Abstract

Rituximab is a chimeric monoclonal antibody which has specific for CD20 antigen expressed by pre-B and mature B-cells. Radiolabelled Rituximab, 131I-Rituximab, has been sucessfully used for treatment of B-Cell NHL. Due to its short shelf-life, 131I-Rituximab is commonly freshly prepared in hospitals prior to its used.  This study aimed to validate rituximab iodination procedure for 131I-Rituximab preparation in order to find the most suitable procedure to be applied in hospitals which intend to produce 131I-Rituximab in-house.  Three different methods of radiolabelling using three types of oxidizing agents, namely Iodobeads, Iodogen, and Chloramine-T were performed. Prior to the validation, radiochemical purity test and purification procedures were also validated as these procedures are critical for producing an acceptable quality of  I-Rituximab. In addition, the shelf-life of 131I-Rituximab was also studied. This study was conducted at the Centre for Radioisotope and Radiopharmaceutical Technology, Serpong during the period of July 2015 to February 2018.  The results showed that the radiochemical purity test of 131I-Rituximab could be easily performed by using instance thin layer chromatography?silica gel (ITLC-SG) in the stationary phase and 85% methanol or saline in the mobile phase. Purification of 131I-Rituximab was conducted using a Sephadex G-25 M filled column with 0.1 M PBS, pH 7.2, as the eluent that was found to be quite reliable to give 131I-Rituximab with radiochemical purity of >95% and recovery of approximately 90%. Radiolabelling efficiency performed using Iodobeads was the lowest (60%) compared to that of Iodogen and Chloramine-T (80?90%). In addition, approximately 30% of  I was retained by Iodobeads and this procedure was time consuming(~ 1 hours). It is concluded that Chloramine-T and Iodogen are better than Iodobeads as the oxidizing agent for radiolabelling of Rituximab with 131I. The radiochemical purity of 131I-Rituximab is well maintained when stored at room temperature and in 4 °C temperature up to 6 hours.Validasi Prosedur Iodinasi Rituximab untuk Preparasi131 I-RituximabValidasi proseduri odinasi rituximab untuk preparasi131I-Rituximab telah berhasil dilakukan.  Validasi ini dilakukan untuk mendapatkan prosedur yang paling sesuai yang dapat diaplikasikan untuk produksi131 I-Rituximabdi rumah sakit yang ingin memproduksi131 I-Rituximab di laboratorium mereka.  Tiga metode radiolabelling menggunakan 3 jenis oksidator Iodobeads, Iodogen, dan Chloramine-T telah divalidasi. Sebelum validasi ini, prosedur uji kemurnian radiokimia dan pemurnian divalidasi terlebih dahulu karena prosedur-prosedur ini sangat berpengaruh dalam penyediaan131 I-Rituximab dengan kualitas yang baik. Disampingitu, lama simpan131I-Rituximab juga dipelajari. Penelitan ini dilaksanakan di Pusat Teknologi Radioisotop dan Radiofarmaka, Serpong, Juli 2015?Februari 2018. Hasil penelitian memperlihatkan bahwa uji kemurnian radiokimia 131I-Rituximab dapat dilakukan dengan mudah menggunakan instance thin layer chromatography ? silica gel (ITLC-SG) sebagai fasa diam dan metanol 85% atau larutan salin sebagai fasa gerak. Pemurnian131I-Rituximab menggunakan kolom Sephadex G-25 M dan0.1 M PBS pH 7,2 sebagai eluen dapat diandalkan dan memberikan131I-Rituximab dengan kemurnian radiokimia >95% dan sekitar 90% perolehan kembali. Efisiensi penandaaan menggunakan Iodobeads didapatkan paling (60%) dibanding dengan Iodogen dan Chloramine-T (80 ? 90%). Di samping itu, sekitar 30% 131I hilang karena terikat pada Iodobeads dan prosedur ini memakan waktu yang panjang (~1 jam). Penandaan Rituximab 131I menggunakan Chloramine-T and Iodogen dapat disimpulkan lebih baik dibanding dengan menggunakan Iodobeads. Kemurnian radiokimia131I-Rituximab terjaga dengan baik pada penyimpanan selama 6 jam pada suhu kamar dan 4 °C.