ABSTRACT

       Background and Objective: Prostate specific antigen (PSA) is considered as one of the most reliable biomarkers of cancer and other known prostate diseases. In the present study, solid phase sandwich immunoradiometric assay was used to measure the amount of PSA. In this type of measurement, a pair of anti-PSA antibodies on the solid phase and labeled with Iodine-125, participate in forming a complex with two different epitopes of PSA.

       Methods: Variables such as irradiation level, modification of polymer surfaces by alcohol washing, different concentrations and volumes of antibody, incubation temperature and drying conditions that influence the direct coating process were optimized. Finally, the stability, accuracy and precision of the laboratory kit were evaluated by comparison with a foreign kit.

      Results: According to the obtained results, preliminary preparations such as irradiation, tube washing and specific temperature conditions are not required during the coating process. Drying by lyophilization method does not affect the quality of coating. Antibody concentration of 2.5 μg/ml and coating volume of 800 μl were determined as the optimum conditions for coating, which had good stability within a year. Alignment of results obtained from the domestic and foreign kits for accuracy of 30 samples from patients was confirmed by T-test (sig 2-tailed= 0.993 and 95% confidence interval). The short-term and long-term precision for three control ranges (low, medium, high) were less than 0.25 and 0.33 of allowable total error (TEa = 10%), respectively.

       Conclusion: The produced domestic kit has acceptable precision according to the CLIA criteria.

       Keywords: Biological testing, Radioimmunometric assay, monoclonal antibody, prostate specific antigen, prostate disease.

ABSTRACT
          Background and Objectives: Poliomyelitis remains a major public health problem in developing countries, which signify the need for extensive diagnostic and prevention research. The aim of the present study was to design monoclonal antibodies (MAbs) against poliovirus type I with biomedical, diagnostic and therapeutic applications.
          Methods: B-cells were isolated from a mouse challenged with polio antigen injection. The B-cell were fused with myeloma tumor cells. After evaluation and screening of approximately 250 hybridoma colons by ELISA, 35 colons with the highest antibody titer and no cross-reactivity were selected and subsequently cloned by limiting dilution. Finally, three colons capable of secreting MAbs against epitopes of poliovirus type I were used for MAb production. Next, the MAbs were characterized by antibody assays, isotyping, epitope analysis (western blot), cross-reactivity test, stability test, sterility test and mycoplasma test.
          Results: The results indicated that the MAbs were of IgG1 kappa chain, had good stability and no cross-reactivity. In western blot, a band at 26 kDa which is associated to VP3 neutralization protein was observed.
          Conclusion: These serotype-specific MAbs can be potentially used for identification of type I poliovirus for research, diagnostic and prevention purposes.
          Keywords: Monoclonal antibody, Hybridoma, Poliomyelitis, Poliovirus.