Profiling human serum apolipoproteins by mass spectrometry /
The construction and performance of an electrospray ionization time-of-flight (ESITOF) mass spectrometer with new features are described. The variables that influence mass measurement resolution, accuracy and sensitivity were studied and optimized using theoretical calculations and experimental adj...
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| Format: | Thesis Book |
| Language: | English |
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[Place of publication not identified] :
[publisher not identified] ;
1998.
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| Online Access: | http://proxy.library.tamu.edu/login?url=http://proquest.umi.com/pqdweb?did=737708161&sid=1&Fmt=2&clientId=2945&RQT=309&VName=PQD |
| Summary: | The construction and performance of an electrospray ionization time-of-flight (ESITOF) mass spectrometer with new features are described. The variables that influence mass measurement resolution, accuracy and sensitivity were studied and optimized using theoretical calculations and experimental adjustments. The linear TOF analyzer incorporates an orthogonal accelerator (OA) with a triple acceleration field. A set of distances and acceleration voltages of the OA was optimized to obtain mass resolution of - 1850 (FWHM). Examples of the application of the system include a study of ion-molecular collisions in the EIG region, mass accuracy, thermally induced dissociation and charge state distributions of several commercial proteins. In the second half of the dissertation, the newly developed OA ESI-TOF mass spectrometer and a commercially available delayed extraction matrix-assisted laser desorption/ionization (DE MALDI-TOF) instrument were employed to characterize natural mixtures of human serum apolipoproteins (apos). The analytical parameters of the two instruments were optimized for accurate ( ︢I Da), precise (SD = - I Da), and high resolution MW measurement of the apos from very low density lipoprotein (VLDL) and high density lipoprotein (HDL) fractions in the molecular weight (MW) range from 6 kDa to 30 kDa. The most important feature of the method is that it is a multi-component analysis with high specificity over the entire isoform distribution and in a single measurement requiring a few minutes of data acquisition time. More than twenty different A and C apo isoforms have been identified in delipidated HDL fraction from a young healthy donor in this study, including several new human serum apo A and apo C isoforms. The discovered isoforms have the same relative charge and similar MW values compare to the mature apo isoforms. This feature precludes their efficient separation and identification by the 2-D gel electrophoresis technique, which has been used to screen for apo polymorphism in the past. The newly developed method opens a new dimension for the human proteom research. |
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| Item Description: | Vita. "Major Subject: Chemistry". |
| Physical Description: | xv, 176 leaves : illustrations ; 28 cm. Issued also on microfiche from University Microfilms Inc. |
| Bibliography: | Includes bibliographical references: pages 147-171. |