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Wening Lestari
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Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Materials Science & Nanotechnology

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OPTIMASI PEMBUATAN COATED TUBE HUMAN SERUM ALBUMIN (HSA) UNTUK KIT RADIOIMMUNOASSAY (RIA) MIKROALBUMINURIA Sutari Sutari; V. Yulianti S.; Triningsih Triningsih; Gina Mondrida; Agus Ariyanto; Sri Setiyowati; Puji Widayati; Wening Lestari
Jurnal Forum Nuklir JFN Vol 8 No 1 Mei 2014
Publisher : BATAN

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (2601.863 KB) | DOI: 10.17146/jfn.2014.8.1.3476

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.
PREPARASI PEREAKSI KIT IMMUNORADIOMETRlCASSAY FREE PROSTATE SPECIFIC ANTIGEN UNTUK DETEKSI KANKER PROSTAT Puji Widayati; Gina Mondrida; Sri Setiyowati; Agus Ariyanto; V. Yulianti Susilo; Wening Lestari
Jurnal Kimia Terapan Indonesia Vol 15, No 2 (2013)
Publisher : Research Center for Chemistry - LIPI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (5744.033 KB) | DOI: 10.14203/jkti.v15i2.107

Abstract

Prostate Specific Antigen (PSA) is a glycoprotein with a molecular weight of approximately 34,000 daltons serine protease secreted exclusively by prostatic epithelial cells that lining acini and prostate gland. Increased of PSA levels can be caused by prostate cancer or benign prostate enlargement (benign prostatic hyperplasia, BPH). PSA in the blood was found in the free condition (free PSA) and most of the bound protein (complexed-PSA, c-PSA). Measuring levels of PSA 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 radionuclides ,/' 125 oJ I as a tracer, so the sample in small 13 quantity can be detected The purpose of this study was obtained PSA reagent kit that includes 1251labeled PSA as a tracer, PSA coated tube and PSA standard that requirements of the kit, then it can be optimized assay design, that eventually PSA reagent kit can be used for early detection of prostate cancer. It has been done labeling of Mab PSA using 125 1with reaction time was 90 seconds, amount of PSA MAb was 75 ugram and the activity of Na_ 125I was 1000 flCi. Preaparation of PSA coated tube using 0.05 M Na2C03 solution, at pH: 9.6 with volume was 250 ml., standard PSA with 0.025 Mphosphate buffer at pH 7.4 containing 5% BSA and 0.1% NaN3, and resulting at 1,25% and 14,12% respectively of NSB and BIT that requirement of the kit.Keywords: Prostate cancer, PSA, IRMA,NSB, Maximum Binding
Co-Authors Agus Ariyanto Gina Mondrida Puji Widayati Puji Widayati Sri Setiyowati Sutari Sutari Triningsih Triningsih V. Yulianti S. V. Yulianti Susilo