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Aufreinigung und Konzentrierung anaboler Steroide mittels der Immunoaffinitätschromatographie

Aufreinigung und Konzentrierung anaboler Steroide mittels der Immunoaffinitätschromatographie

          
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About the Book

Inhaltsangabe: Einleitung: Die in der Spurenanalytik von pharmakologisch wirksamen Substanzen eingesetzten Extraktionsverfahren reichen oft nicht aus, um die gesuchte Substanz in der biologischen Matrix (Urin, Blut, Haar, etc.) eindeutig nachzuweisen. Das liegt zum einen daran, daß die Konzentrationen dieser Substanzen äußerst gering sind und zum anderen, daß die Bioprobe eine solch komplexe Matrix bildet, daß sich Interferenzen körpereigener Stoffe bei der anschließenden gaschromatographisch-massenspektrometrischen Detektion nicht vermeiden lassen. Durch eine besondere Aufreinigung, die Immunoaffinitätschromatographie genannt wird, sollen diese Störsubstanzen entfernt werden. Dabei macht man sich das in der Natur einzigartige Prinzip zunutze, nach dem Antikörper ihr korrespondierendes Antigen (zu analysierende Substanz) mit hoher Selektivität erkennen und reversibel binden. Die Antikörper werden auf eine Festphase immobilisiert und anschließend mit der Probe inkubiert. Die gesuchte Substanz kann so nach dem Schlüssel-Schloß-Prinzip von den Antikörpern gebunden werden, während unspezifische Moleküle in der Bioprobe verbleiben. Nach einer Waschphase wird mittels eines organischen Solvens die Dissoziation des Antikörper-Antigen-Komplexes bewirkt und der Analyt in gereinigter Form gewonnen. Die Herstellung von Antikörpern mit der gewünschten Spezifität (z.B. gegen anabole Steroide) wurde in dieser Arbeit durch Kopplung von Steroidhormonen an ein Protein und Immunisierung von Kaninchen mit diesen Steroid-Protein-Konjugaten ermöglicht. Kleine Moleküle in der Größenordnung von Steroiden lösen per se keine Immunantwort und damit keine Produktion von Antikörpern aus. Erst durch die Bindung an ein großes Carriermolekül, wie z.B. Bovine Serum Albumin (BSA) oder Keyhole Limpet Hemocyanin (KLH), werden sie vom Organismus als Fremdsubstanz erkannt und lösen die Antikörperproduktion aus. Die Gruppe der 17alpha-methylierten Steroide zählt zu den am häufigsten mißbräuchlich e


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Product Details
  • ISBN-13: 9783838624662
  • Publisher: Diplom.de
  • Binding: Paperback
  • Language: German
  • Returnable: N
  • Spine Width: 10 mm
  • Width: 148 mm
  • ISBN-10: 3838624661
  • Publisher Date: 02 Jul 2000
  • Height: 210 mm
  • No of Pages: 180
  • Series Title: German
  • Weight: 245 gr

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