Affinity capture mass spectrometry of biomarker proteins using peptide ligands from biopanning.
Affinity capture mass spectrometry was used to isolate and ionize protein A from Staphylococcus aureus from both a commercial source and cell culture lysate using matrix assisted laser desorption/ionization (MALDI) mass spectrometry. Two surfaces are compared: gold surfaces with immunoglobulin G covalently immobilized and silica surfaces with a covalently bound small peptide discovered via biopanning. A detection limit of 2.22 bacterial cells/mL of culture fluid was determined for the immobilized peptide surfaces. This study emphasizes the ability to use peptide ligands to effectively capture a biomarker protein out of a complex mixture. This demonstrates the potential to use biopanning to generate capture ligands for a large variety of target proteins and subsequently detect the captured protein using MALDI mass spectrometry.
Johnson
Erin M
EM
Departments of Chemistry and Biochemistry, University of Arizona, Tucson, Arizona 85721, USA.
Ellis
Walther R
WR
Powers
Linda S
LS
Wysocki
Vicki H
VH
D015415:Biomarkers / Q000032:analysis* / Q000378:metabolism*; D006046:Gold / Q000737:chemistry; D007118:Immunoassay; D007074:Immunoglobulin G / Q000276:immunology / Q000378:metabolism; D010446:Peptide Fragments / Q000276:immunology / Q000378:metabolism*; D012834:Silver / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013205:Staphylococcal Protein A / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D013211:Staphylococcus aureus / Q000254:growth & development / Q000378:metabolism
D015415:Biomarkers; D007074:Immunoglobulin G; D010446:Peptide Fragments; D013205:Staphylococcal Protein A; D012834:Silver; D006046:Gold