The Core Hemoglobin Laboratory is an essential component of the NCCSCC. During the current grant period, (1993-1998), the Core performed standard hemoglobin diagnostic procedures, DNA diagnostic procedures, developed a number of advanced mass spectrometry based techniques for structural analysis of hemoglobin and allied all of these techniques to the analysis of unusual hemoglobin variants. As a result of this work, the Core Hemoglobin Laboratory has become recognized as a national resources for analysis of hemoglobin variants. In the current application, the Core Hemoglobin Laboratory will continue to provide routine hemoglobin diagnostic services, such as TLIF, electrophoresis, and quantitative A2 and F measurements for basic and clinical research projects within the Center. DNA diagnostic studies will be available through clinical laboratories at UCSF and CHORI. Specialized methods available in the Core Hemoglobin Laboratory will be used for diagnosis of rate hemoglobin variants, clinical research-demonstration projects and basic research projects. Diagnostic services will be made available to Sickle Cell Center and Newborn screening programs in cases where clinically symptomatic patients with unknown hemoglobin variants are identified. The Core Hemoglobin Laboratory will participate in basic research projects involving transgenic mice. Mass spectrometric and HPLC techniques will be used to characterized globin expression and constitution of hemoglobin tetramers during development. Globin biosynthetic studies and detection of chimerism in newborn mice will be performed to facilitate the choice of potential breeders. The Core will serve as a quality control resource by evaluating faithfulness of expression of the transgene products and will perform characterization of unexpected hemoglobin protein species in transgenic animals, should they occur. In addition to its commitment to clinical and basic research projects, the Core Hemoglobin Laboratory will undertake studies in method development directed towards microscale analysis of hemoglobins.
The aim of this research will be to develop methodologies involving nanospray electrospray ionization mass spectrometry (ESMS) interfaced with capillary HPLC for structural characterization of hemoglobins whose quantity is very limited (transgenic animal embryos, red cell progenitors and minor hemoglobin components isolated by TLIF).
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