Phage display for identification of serum biomarkers of traumatic brain injury

• Published in: Journal of neuroscience methods Vol. 272; pp. 33 - 37
• Main Authors: Ghoshal, Sarbani, Bondada, Vimala, Saatman, Kathryn E., Guttmann, Rodney P., Geddes, James W.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15.10.2016
• Subjects: Amino Acid Sequence; Animal models; Animals; Bacteriophages - genetics; Bacteriophages - metabolism; Biomarkers; Biomarkers - blood; Blood; Blood Chemical Analysis - methods; Brain Injuries, Traumatic - blood; Brain Injuries, Traumatic - diagnosis; Capsid Proteins - genetics; Capsid Proteins - metabolism; Cell Surface Display Techniques; Concussion; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Glial Fibrillary Acidic Protein - blood; Male; Mice, Inbred C57BL; Parietal Lobe; Peptide Library; Rodents; Traumatic brain injury; Biomarkers; Animal models; Traumatic brain injury; Concussion; Rodents; Blood
• ISSN: 0165-0270; 1872-678X
• DOI: 10.1016/j.jneumeth.2016.04.026

•Phage display is an unbiased approach to identify serum biomarkers.•Applicable to acute CNS injuries as well as neurological disorders.•Identification of GFAP as serum biomarker of TBI provides proof-of-concept. The extent and severity of traumatic brain injuries (TBIs) can be difficult to determine with current diagnostic methods. To address this, there has been increased interest in developing biomarkers to assist in the diagnosis, determination of injury severity, evaluation of recovery and therapeutic efficacy, and prediction of outcomes. Several promising serum TBI biomarkers have been identified using hypothesis-driven approaches, largely examining proteins that are abundant in neurons and non-neural cells in the CNS. An unbiased approach, phage display, was used to identify serum TBI biomarkers. In this proof-of-concept study, mice received a TBI using the controlled cortical impact model of TBI (1mm injury depth, 3.5m/s velocity) and phage display was utilized to identify putative serum biomarkers at 6h postinjury. An engineered phage which preferentially bound to injured serum was sequenced to identify the 12-mer ‘recognizer’ peptide expressed on the coat protein. Following synthesis of the recognizer peptide, pull down, and mass spectrometry analysis, the target protein was identified as glial fibrillary acidic protein (GFAP). GFAP has previously been identified as a promising TBI biomarker. The results provide proof of concept regarding the ability of phage display to identify TBI serum biomarkers. This methodology is currently being applied to serum biomarkers of mild TBI.