Siti Darwati
Pusat Pengembangan Radioisotop dan Radiofarmaka, BATAN, Serpong.

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Journal : Jurnal Radioisotop dan Radiofarmaka

OPTIMALISASI PEMBUATAN KIT IRMA CA-125 Ariyanto, Agus; Darwati, Siti; Mondrida, Gina; Yunita, Fitri; Widayati, Puji; Setiyowati, Sri; Sulaiman, Sulaiman; Yulianti, Veronika; Triningsih, Triningsih
Jurnal Radioisotop dan Radiofarmaka Vol 6, No 2 (2003): Jurnal PRR 2003
Publisher : Jurnal Radioisotop dan Radiofarmaka

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Abstract

ABSTRAK OPTIMALISASI PEMBUATAN KIT IRMA CA-125. Telah dilakukan penelitian optimalisasi pembuatan kit  IRMA  CA-125 yang meliputi pembuatan perunut monoklonal anti CA-125 bertanda 125I dan  pembuatan  "coated tube". Pada pembuatan perunut digunakan 2 macam oksidator yang divariasikan jumlahnya, yaitu kloramin-T dan N- bromosuksinimid.  Optimalisasi pembuatan "coated tube" dilakukan dengan memvariasikan volume larutan "coating", pengaruh pencucian, pengaruh penambahan BSA, pengaruh   bufer  pelarut dan uji pemasangan monoklo-nal anti CA-l25. Hasil percobaan menunjukkan bahwa oksidator  kloramin-T dapat digunakan untuk pembuatan perunut mononoklonl  anti CA-l25 dengan rendemen cukup tinggi yaitu 96.5% dan kemumian radiokimia 93,2%. Jumlah oksidator yang optimal adalah 10 µg. Pembuatan "coated tube  untuk  monoklonal anti CA-l25 dengan menggunakan volume larutan "coating" yang bervariasi antara 300 µI sampai dengan 600 uL tidak menunjukkan perbedaan nyata pada nilai %NSB dan %BT. Proses pencuci "coated tube" menggunakan BSA 0, I % memberikan pengaruh nyata pada %BT. walaupun tidak berpengaruh terhadap nilai %NSB. Penggunaan bufer bikarbonat untuk pembuatan "coated tube" memberikan hasil yang lebih baik dari pada bufer fosfat. Untuk pemasangan monoklonal anti CA-125 diketahui bahwa pasangan monoklonal anti CA-125. monoklonal anti CA-125  M37203M sebagai  perunut dan monoklonal anti CA-125  M86924M sebagai "coating solution" memberikan hasil yang optimal. Kata kunci : Kit IRMA, Monoklonal anti CAll25, Pemnut ABSTRACTOTIMALIZATION FOR PREPf\RATION OF CA-125 IRMA-KIT. Optimalization on thepreparation of CA-125 IRMA-kit has been studied involving preparation of 125I  labeled monoclonalanti CA-125 as a tracer and preparation of coated tubes. Chloramine-T and N-bromosuccinimide were used as oxidators by varying them in their quantity. Optimalization on preparation of coated tube was carried out by variation in volume of  coating solution, effect of washing, addition of BSA 0.1%, buffer and pairing test for the monoclonal anti CA-125. The use of chloramine-T as oxidator for 125I labeling of monoclonal anti CA-15 was found to giv labeling-yield of about 96.5% and radiochemical purity of 93.2%. Optiimum amount of the oxidizing agent was found to be 10µg. There were not any significant difference in utilization of coating solution volume ranging at 300 to 600µL. for preparing monoclonal anti CA-125 coated tube. The non specific binding (NSB) and the %B/T values were found to be unaffected  by washing step, while a blocking of coated tube using of BSA 0.1% decreased the %B/T value. Utilization of bicarbonate buffer for the preparation  of coated tube was found to be more satisfactory than that of phosphate buffer. It was shown that the monoclonal anti CA-125 M37203 used for tracer being paired with monoclonal anti CA-125 M866924M used for coating solution gave an optimal result based on the values of %NSB  and %B/T. Key word : IRMA-Kit, Monoclonal anti CA-125, Tracer.   
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