نوع مقاله : مقاله پژوهشی
نویسندگان
مرکز تحقیقات هسته‌ای، سازمان انرژی اتمی، صندوق پستی: 3486-11365 تهران- ایران
چکیده
روش‌های تعیین غلظت پادگن ویژه پروستات (PSA) در خون که شاخصی انتخابی برای تشخیص سرطان پروستات و پیگیری درمان آنند, اغلب بر اساس سنجش ایمن‌شناختی خاص پایه‌گذاری شده‌اند؛ در این روش‌ها از ایجاد کمپلکس پادگن- پادتن برای تعیین مقدار PSA استفاده می‌شود. با انجام دادن این تحقیق, پادگن PSA به مقدار کافی بصورت نیمه‌خالص و خالص تهیه شد که ماده‌ای ارزشمند در تهیه استاندارد برای کیت‌های تشخیصی PSA می‌باشد؛ سپس با استفاده از این پادگن, هشت نوع پادتن تک‌تاکی (مونوکلونال) علیه PSA بدست آمد که شامل 4 نوع پادتن تک‌تاکی زوج هستند و می‌توان از هر زوج برای تهیه یک کیت IRMA (کیت ساندویچی) استفاده کرد. با انجام دادن آزمایش تعیین میزان تمایل (affinity) پادتن به پادگن, از میان این چهار زوج, یک زوج با تمایل زیاد برای تهیه کیت IRMA بکار برده شد. این کیت از نمودار استاندارد مطلوبی برخوردار بوده و قادر است غلظت PSA در سرم را تا 0.1نانوگرم در میلی‌لیتر اندازه‌گیری نماید.
کلیدواژه‌ها
عنوان مقاله [English]
Development of a New in Vivo Kit for Detection of Prostate Specific Antigen in Human Serum Using Immunoradiometric Assay Method
نویسندگان [English]
- M.H Babaei
- P Behradkia
- M Shafii
- M Moula
- H Forutan
- R Najafi
چکیده [English]
Prostate is a leading site for the cancer incidence, accounted for 31.0% of new cancer cases in men. Prostate-specific antigen (PSA) is widely used in the detection and monitoring of the prostate cancer. Currently, immunoassay is used to detect PSA in human serum. This technique is based on the interaction between antibody and antigen. The varied immunoassay formats and equipment to run the assays allow the users to measure the analytes rapidly, with the flexibility to run a small or a large number of samples. Among different immunoassay methods, immunoradiometric assay (IRMA) is a more sensitive and valuable detection approach. This study has been made in 4 parts: (1) purification of PSA from seminal fluid; (2) preparation of hybridoma cells which secrete monoclonal antibody (mAb) against PSA, (3) selection of pair monoclonal antibody among those antibodies, and finally (4) design of an IRMA kit and it’s quality control . The results of this study were: (1) obtaing a huge amount of PSA as semi-purified and purified, that is a valuable material for preparation of standard kits; (2) preparation of 8 kinds of monoclonal antibodies; (3) finding 4 pairs of monoclonal antibodies which react with different epitopes on PSA molecule; and (4) preparation of IRMA kit for measuring PSA concentration in human serum.
کلیدواژه‌ها [English]
- Prostate Specific Antigen (PSA)
- Immunoradiometric Assay (IRMA)
- Prostate Cancer
- Screening test
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