A novel recombinant antibody specific to full-length stromal derived factor-1 for potential application in biomarker studies

• Published in: PloS one Vol. 12; no. 4; p. e0174447
• Main Authors: Bromage, Daniel I, Taferner, Stasa, Pillai, Mahesh, Yellon, Derek M, Davidson, Sean M
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 05.04.2017; Public Library of Science (PLoS)
• Subjects: Adolescents; Amino acids; Anesthesia; Angina; Animals; Antibodies; Antibodies - immunology; Binding sites; Biology and Life Sciences; Biomarkers; Biomarkers - blood; Biosensors; Blood; Blotting, Western; c-Kit protein; Cardiac conditioning; Cardiovascular diseases; Cathepsin G; CD34 antigen; Cell adhesion; Cell adhesion & migration; Cerebrospinal fluid; Chemokine CXCL12 - blood; Chemokine CXCL12 - immunology; Chemokines; Circadian rhythms; Diabetes mellitus; Dipeptidyl Peptidase 4 - blood; Dipeptidyl-peptidase IV; Enzyme-Linked Immunosorbent Assay; Ethics; Genetic aspects; Health risks; Heart attacks; Heart diseases; Heart failure; Hypoxia; Hypoxia - blood; Hypoxia - diagnosis; Hypoxia - metabolism; Immunoglobulin G; Immunoglobulins; Inhibition; Ischemia; Ischemia - blood; Ischemia - diagnosis; Ischemia - metabolism; Isoforms; Kinases; Kinetics; Lymphocytes; Male; Mass spectrometry; Medical screening; Medicine and Health Sciences; Myocardial infarction; N-Terminus; Packaging; Peptides; Proteins; R&D; Rats; Rats, Sprague-Dawley; Recombinant Proteins; Research & development; Research and Analysis Methods; Risk factors; Rodents; Sensors; Sodium; Splicing; Stem cells; Studies; Surgery; Transplantation; Ventricle
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0174447

Stromal derived factor-1α (SDF-1α/CXCL12) is a chemokine that is up-regulated in diseases characterised by tissue hypoxia, including myocardial infarction, ischaemic cardiomyopathy and remote ischaemic conditioning (RIC), a technique of cyclical, non-injurious ischaemia applied remote from the heart that protects the heat from lethal ischaemia-reperfusion injury. Accordingly, there is considerable interest in SDF-1α as a potential biomarker of such conditions. However, SDF-1α is rapidly degraded and inactivated by dipeptidyl peptidase 4 and other peptidases, and the kinetics of intact SDF-1α remain unknown. To facilitate investigation of full-length SDF-1α we established an ELISA using a novel recombinant human antibody we developed called HCI.SDF1. HCI.SDF1 is specific to the N-terminal sequence of all isoforms of SDF-1 and has a comparable KD to commercially available antibodies. Together with a detection antibody specific to the α-isoform, HCI.SDF1 was used to specifically quantify full-length SDF-1α in blood for the first time. Using RIC applied to the hind limb of Sprague-Dawley rats or the arms of healthy human volunteers, we demonstrate an increase in SDF-1α using a commercially available antibody, as previously reported, but an unexpected decrease in full-length SDF-1α after RIC in both species. We report for the first time the development of a novel recombinant antibody specific to full-length SDF-1. Applied to RIC, we demonstrate a significant decrease in SDF-1α that is at odds with the literature and suggests a need to investigate the kinetics of full-length SDF-1α in conditions characterised by tissue hypoxia.