Immunoproteomics.
A novel immunoproteomic assay, combining specificity of antibody with precision of mass spectral analysis is described, and a number of practical applications are presented. The assay is carried out in three steps. The first step of the assay involves antibody immobilization, using a bacterial Fc binding support. The second step is antigen capture and washing to remove non-specific binding. The third step involves analysis of the captured antigens by SELDI-TOF. The assay has many advantages in sensitivity, speed, and economy of reagents in detection of specific antigens or antibodies. In addition, under appropriate experimental conditions, semi-quantitative data may be obtained. By combining the increasing range of selective specific antibody reagents available, in part due to advances in antibody engineering technology, and the resolving power available, using mass spectrometry, immunoproteomics is a valuable technique in proteomic analysis. A number of examples of the application of this technique to analysis of biological systems are presented.
Hess
Jennifer L
JL
Department of Biology, Juniata College, 1700 Moore Street, Huntingdon, PA 16652-2196, USA.
Blazer
Levi
L
Romer
Terence
T
Faber
Lee
L
Buller
R Mark
RM
Boyle
Michael D P
MD
D000818:Animals; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D000941:Antigens / Q000032:analysis*; D006801:Humans; D007141:Immunoglobulin Fc Fragments; D007371:Interferon-gamma / Q000032:analysis; D013058:Mass Spectrometry / Q000379:methods; D040081:Protein Array Analysis; D011506:Proteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D012709:Serum Albumin / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000941:Antigens; D007141:Immunoglobulin Fc Fragments; D011506:Proteins; D012709:Serum Albumin; D007371:Interferon-gamma