An Immunoproteomic Approach for Identification of Clinical Biomarkers for Monitoring Disease

Abstract

Circulating antibodies can be used to probe protein arrays of body fluids, prepared by two-dimensional gel electrophoresis, for antigenic biomarker detection. However, detected proteins, particularly low abundance antigens, often remain unidentifiable due to proteome complexity and limiting sample amounts. Using a novel enrichment approach exploiting patient antibodies for isolation of antigenic biomarkers, we demonstrate how immunoproteomic strategies can accelerate biomarker discovery. Application of this approach as a means of identifying biomarkers was demonstrated for cystic fibrosis (CF) lung disease by isolation and identification of inflammatory-associated autoantigens, including myeloperoxidase and calgranulin B from sputum of subjects with CF. The approach was also exploited for isolation of proteins expressed by the Pseudomonas aeruginosa strain PA01. Capture of PA01 antigens antibodies from subjects of Pseudomonas subjects Pseudomonas proteins, and protein and proteins, as biomarkers of Circulating antibodies can be used to probe protein arrays of body fluids, prepared by two-dimensional gel electrophoresis, for antigenic biomarker detection. However, detected proteins, particularly low abundance antigens, often remain unidentifiable due to proteome complexity and limiting sample amounts. Using a novel enrichment approach exploiting patient antibodies for isolation of antigenic biomarkers, we demonstrate how immunoproteomic strategies can accelerate biomarker discovery. Application of this approach as a means of identifying biomarkers was demonstrated for cystic fibrosis (CF) lung disease by isolation and identification of inflammatory-associated autoantigens, including myeloperoxidase and calgranulin B from sputum of subjects with CF. The approach was also exploited for isolation of proteins expressed by the Pseudomonas aeruginosa strain PA01. Capture of PA01 antigens antibodies from subjects of Pseudomonas subjects Pseudomonas proteins, and protein and proteins, as biomarkers of due to protein of expressed proteins protein as a of disease disease biomarker and can be used two-dimensional gel cystic used two-dimensional gel cystic to the and of to biomarker body fluids, as can be by of biomarkers, particularly low abundance proteins, by protein of body the of protein with a of sample including of abundance proteins of protein biomarkers proteome sample with the of the to low abundance and of from subjects as and a means for antigenic proteins protein from body fluids, be of identification of protein as a a lung gel as for identifying of a identification and of of antigenic proteins immunoproteomic of antigens remain unidentifiable due to low abundance by the of and and of lung of to biomarker exploited the of antibodies from of protein for and and and of antigenic for enrichment for and identification of proteins protein to biomarker discovery. antigenic biomarker this and identification of antigens be of low abundance and of to biomarker biomarker from of the disease and used to antigens from as sputum antigens by to protein for of including identification and we demonstrate how protein can be used for of protein a and protein demonstrate of immunoproteomic to a of antigenic biomarkers for cystic fibrosis fibrosis disease by the cystic fibrosis of the cystic fibrosis and of of from and for to due to of particularly Pseudomonas aeruginosa and and of cystic to lung the of and the of biomarkers of the of subjects by to and to lung as a of of and from aeruginosa PA01 was from and by sputum was and as of by sputum and from and the of from with and from with a cystic of disease cystic and aeruginosa subjects the of subjects also a of to the subjects of the of of protein from sputum was used for of was used for aeruginosa protein and to sputum by and protein and protein and protein with a of of protein of 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