İmmünolojik tanı kitlerinin geliştirilmesi
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- Tez No: 355106
- Danışmanlar: PROF. DR. BEYAZIT ÇIRAKOĞLU
- Tez Türü: Doktora
- Konular: Tıbbi Biyoloji, Medical Biology
- Anahtar Kelimeler: Belirtilmemiş.
- Yıl: 1997
- Dil: Türkçe
- Üniversite: Marmara Üniversitesi
- Enstitü: Sağlık Bilimleri Enstitüsü
- Ana Bilim Dalı: Tıbbi Biyoloji Ana Bilim Dalı
- Bilim Dalı: Belirtilmemiş.
- Sayfa Sayısı: 150
Özet
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Özet (Çeviri)
It is necessary to determine some of the proteins and hormons which exist in the body fluids such as urin, serum or mucus for diagnosis of some disease. In order to determine these proteins and hormons (antigens) by using immunological methods, some antibodies which are binding to them are required. In this study, clinically important antigens and some monoclonal and polyclonal antibodies developed against these antigens were choiced. Some of the antigens were purified and test systems which are based on antigen-antibody interactions were developed. Using hybridoma technology, some hybrid cells which produce anti-HbA and anti-Transferrin monoclonal antibodies were produced. Monoclonal antibodies from supernatant of culturated hybrid cell and prepared antybody - enzyme conjugates were prufied by using various protein prufication methods. The application of the immunological test systems are based on antibodies. These systems are different depending on the structure of the antibody (monoclonal or polyclonal) and body fluids. We used generally indirect, competitive or sandwich ELISA systems. We developed enzyme-linked immunoassays using monoclonal or polyclonal antibodies against fhM (~2 Microglobulin) or hCG (Human Chorionic Gonadotropin). We prepared second antibody-enzyme conjugates m order to use in immunological kits. Polyclonal antibodi:::s developed in rabbits against mouse antibodies. Then these antibodies were orufied from antiserum and conjugated with AP. After conjugation with AP of anti-rabbit antibodies they are used in the kit system with policlonal rabbit antibodies. Hybrid c~lls are produced from culture medium withoutserum. Using antitransferrin monoclonal antibodies we determined that the transferrin is an important component of culture medium without serum. It was also showed that in culture medium without serum, the epitop of the anti-transferrin is in competition with transferrin in serum or urin. The epitop effected negatively the cell production, and the relationship with its transferrin receptor. The stability and activity of immunological diagnosis kits were investigated at its different time intervals.
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