Document Type : Research Articles
Authors
1
Doctoral Program in Medical Science, Faculty of Medicine, Universitas Brawijaya, Malang, Indonesia.
2
Urology Department, Faculty of Medicine, Universitas Brawijaya, Saiful Anwar Hospital, Malang,
3
Department of Biochemistry and Biomolecular, Faculty of Medicine, Universitas Brawijaya, Malang, Indonesia.
4
Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Brawijaya, Malang, Indonesia.
5
Department of Clinical Microbiology, Faculty of Medicine, Universitas Brawijaya, Malang, Indonesia.
Abstract
Objective: This study will identify specific epitopes from the 47kDa protein as the basis for making polyclonal antibodies to increasing sensitivity and specificity of 47kDa protein as bladder cancer biomarkers. Method: The 47kDa protein epitope prediction was carried out using the in-silico method. The epitope with the highest and the lowest value was immunized to the mice for four weeks and was harvested at the fifth weeks. The antibody was tested with the patient’s urine using western blotting. Total of 186 participants including in this study. For the first stage (antibody confirmation test) test we have 18 participants, for the second stage (1st antibody diagnosis test) we have 72 participants and for the third stage (2nd antibody diagnosis test) we have 96 participants, consist of total 64 BC patients 48 of healthy individuals and 74 participants with the other diseases. Results: Some epitopes from the sequenced protein are candidates for immunization, in the chain 108’-136’ (with lowest Bepipred score: 0.53) named as peptide1 and chain 42’-56’ (with highest bepipred score: 0.58) named as peptide2. In western blotting test, both antibodies showed detection at 47kDa. When examined with western blot using urine from BC patients, urine from other cancer patients (prostate, kidney, ureter, rectal, breast), and healthy persons, both antibodies were found to only express 47kDa in urine from BC patients. The diagnostic tests showed high sensitivity (91.67%) and specificity (94.44%) inAb2 in predicting bladder cancer. Conclussion: The evolution of the polyclonal antibody made from specific epitopes is proven to express specifically on bladder cancer patients and have high sensitivity and specificity to diagnose bladder cancer.
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