Analytical Services

Analysis of drug levels in biological fluids (whole blood, plasma, urine)

• Drug and metabolites are extracted from biological specimens using liquid liquid or solid phase extraction. Levels are measured using high-performance liquid chromatography (HPLC) or liquid chromatography/mass spectrometry/mass spectrometry (LC/MS/MS).

Biochemical Services

Transportation of Samples

• Samples are transported from the operating room, General Clinical Research Center or outpatient clinic to the Facility. Tissues are logged into a database and stored at -80°C. Blood is handled immediately upon arrival according to the particular protocol (lymphocyte isolation or DNA isolation) and stored.
  
  Note: The Biochemical Core itself does not actually provide the transportation through any of its direct staff members, though transportation is provided by UChicago messengers.

Plasma Isolation

• Plasma is isolated from whole blood by centrifugation and stored at -80°C until further analysis. In addition, plasma can be centrifuged through Amicon Centrifree Filters to obtain the ultra-filtrate.

Lymphocyte Isolation

• Lymphocytes are isolated from whole blood by centrifugation over a Histopaque (Sigma) gradient or using a CPT Vacutainers. Cells are washed twice, red blood cells are lysed, and pellets are stored at -80°C until further analysis.

DNA Isolation

• DNA is isolated using a Qiagen kit (Qiagen DNeasy Tissue Kit for tissue or cultured cell samples and Qiagen QIAamp DNA Blood Kit for blood samples) and the resulting DNA suspension is stored frozen at -80°C.

RNA Isolation

• Total RNA is isolated using the Qiagen RNeasy Kit, and the resulting RNA suspension is stored frozen at -80°C.

Assessment of Markers for Drug Resistance

• O6-alkylguanine-DNA alkyltransferase (AGT) activity is measured for various clinical trials.