The specific aim of the proposed research is to continue to develop the analytical capability of 252Cf-plasma desorption mass spectrometry for biomedical research involving large biomolecules (m/z greater than 5000). Part of the plan is to change the method of sample preparation so that the formation of molecular ions from the substrate will be less dependent on matrix effects and at the same time will improve sensitivity to the picomole level. The new method of sample preparation will utilize adsorption of proteins (insulin and cytochrome-C) onto polymeric surfaces from liquid solutions. The interaction will be analyzed using phase equilibrium thermodynamics to evaluate equilibrium constants, DeltaH, and DeltaS for adsorption. These same surfaces will become samples for 252Cf-PDMS analysis. Fractional coverage will be determined by 252Cf-PDMS. A general study of polymer-protein surface thermodynamics will be carried out using 252Cf-PDMS as an analytical method. Two important protein systems, interferon and hemoglobin, have been targeted as the next model compounds for extending the mass range and capabilities of 252Cf-PDMS. Some feasibility studies will be carried out to determine whether 252Cf-PDMS might be used to study antibody-antigen interactions and receptor sites on cell surfaces. The studies of protein-polymer interactions have relevance to the genesis of atherosclerosis. Specific interactions between proteins and receptor sites are fundamental to the immune response in humans and in understanding the role of proteins such as insulin in triggering a response within a cell when it attaches to a receptor site.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM026096-10
Application #
3273563
Study Section
Metallobiochemistry Study Section (BMT)
Project Start
1979-07-01
Project End
1988-06-30
Budget Start
1987-07-01
Budget End
1988-06-30
Support Year
10
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Texas A&M University
Department
Type
Schools of Arts and Sciences
DUNS #
City
College Station
State
TX
Country
United States
Zip Code
77845
Song, Siqing; Macfarlane, Ronald D (2002) PDMS-chemistry of angiotensin II and insulin in glucose glass thin films. Anal Bioanal Chem 373:647-55
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Cruzado, I D; Cockrill, S L; McNeal, C J et al. (1998) Characterization and quantitation of apolipoprotein B-100 by capillary electrophoresis. J Lipid Res 39:205-17
Macfarlane, R D; Bondarenko, P V; Cockrill, S L et al. (1997) Development of a lipoprotein profile using capillary electrophoresis and mass spectrometry. Electrophoresis 18:1796-806
Cruzado, I D; Song, S; Crouse, S F et al. (1996) Characterization and quantitation of the apoproteins of high-density lipoprotein by capillary electrophoresis. Anal Biochem 243:100-9
Cruzado, I D; Hu, A Z; Macfarlane, R D (1996) Influence of dodecyl sulfate ions on the electrophoretic mobilities of lipoprotein particles measured by HPCE. J Capillary Electrophor 3:25-9
Bunk, D M; Macfarlane, R D (1992) Fragmentation of proteins in the 13- to 29-kDa mass range observed by 252Cf-plasma desorption mass spectrometry. Proc Natl Acad Sci U S A 89:6215-9
Macfarlane, R D (1990) Principles of californium-252 plasma desorption mass spectrometry applied to protein analysis. Methods Enzymol 193:263-80
Jardine, I; Hunter, S W; Brennan, P J et al. (1986) Heterogeneity of bacterial antigenic lipooligosaccharides determined by californium-252 plasma desorption mass spectrometry. Biomed Environ Mass Spectrom 13:273-6
McNeal, C J; Macfarlane, R D; Jardine, I (1986) A novel mass spectrometric procedure to rapidly determine the partial structure of heparin fragments. Biochem Biophys Res Commun 139:18-24

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