Polyclonal antibody from 47 kDa protein of bladder cancer is sensitive and specific for detection of bladder cancer

Abstract

This study aims to investigate whether the protein isolated from bladder cancer in an Indonesian population can produce the polyclonal antibody for clinical markers of bladder cancer. The participants in this study are bladder cancer patients and healthy persons who were approved for midstream portion urine collection. The inclusion criteria included bladder cancer patients who obtained hematuria examination, urine cytology, and initial therapy with transurethral resection of the bladder; a healthy volunteer who was approved for midstream portion urine collection but without hematuria history, bladder stones, and signs or symptoms of tractus urinarius infection. The procedure consisted of tissue preparation, tissue processing, isolation of membrane cell protein, monitoring of protein in membrane cell, production of polyclonal antibodies, and dot blot technique. A protein with 122 kDa molecular weight is present in epithelial cells of bladder cancer and the normal bladder. A protein with 69 kDa molecular weight is only present in the epithelial cell bladder of normal individuals. In addition, a protein with molecular weight of 47 kDa is only present in epithelial cells of bladder cancer. The minimal ratio of polyclonal antibody with antigen is 1:6400 of the antibody and 1:40 of antigen. Subsequently, this concentration was applied to detect proteins with 47 kDa only in several cancer tissues. Positive results in bladder cancer, but negative results in prostate cancer, rectal cancer, and breast cancer were found. The polyclonal antibody produced from 47 kDa protein subunit is sensitive and specific to detect bladder cancer and become an alternative biomarker for diagnosis and surveillance of bladder cancer.