SIVI BİYOPSİ YAKLAŞIMINDA ÜRİNER EKSTRASELÜLER VEZİKÜLLER: BİYOGENEZ, İZOLASYON, KARAKTERİZASYON VE BİYOKİMYASAL ANALİZLER
Özet
Üriner ekstraselüler veziküller (uEV’ler), böbrek ve üriner sistem hücrelerinden idrara salınan, lipit çift tabaka ile çevrili parçacıklardır. Protein, nükleik asit, lipit ve metabolit kargoları nedeniyle hücresel kökenleri ve biyokimyasal durum hakkında bilgi sağlayabilirler. Bu özellikleri, uEV’leri sıvı biyopsi araştırmaları için önemli hale getirmiştir.
Bu bölümde uEV’lerin biyogenezi, idrara salınımı ve biyokimyasal bileşimi ele alınmıştır. Örnek toplama, ilk işleme, saklama ve dondurma-çözme koşulları gibi preanalitik değişkenler değerlendirilmiştir. Uromodulin, albumin ve diğer kontaminantların EV geri kazanımı ve analiz sonuçları üzerindeki etkileri açıklanmıştır. Diferansiyel ultrasantrifüj, ultrafiltrasyon, boyut dışlama kromatografisi, polimer temelli presipitasyon, immünoaffinite ve mikroakışkan sistemler karşılaştırılmıştır. Parçacık sayısı, boyut dağılımı, morfoloji ve protein belirteçlerine dayalı karakterizasyon yöntemleri özetlenmiştir. Ayrıca Western blot, ELISA, LC-MS/MS, PCR, dizileme, lipidomik ve metabolomik yaklaşımların analitik özellikleri incelenmiştir.
Hiçbir izolasyon yöntemi bütün EV alt gruplarını tam saflıkta elde edemediğinden, sonuçlar kullanılan yöntem bağlamında yorumlanmalıdır. Güvenilir uEV analizleri için tamamlayıcı karakterizasyon yöntemleri kullanılmalı, uygun normalizasyon stratejileri belirlenmeli ve analitik validasyon izolasyondan son ölçüme kadar bütün iş akışını kapsamalıdır.
Anahtar Kelimeler: Üriner ekstraselüler veziküller, sıvı biyopsi, izolasyon, karakterizasyon, biyokimyasal analiz, analitik validasyon
Konu Alanı: Tıp -> Tıp (diğer) -> Biyokimya (tıbbi)
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