Bioaffinity-based surface immobilization of antibodies to capture endothelial colony-forming cells

• Published in: PloS one Vol. 17; no. 8; p. e0269316
• Main Authors: Boulanger, Mariève D, Level, Hugo A, Elkhodiry, Mohamed A, Bashth, Omar S, Chevallier, Pascale, Laroche, Gaétan, Hoesli, Corinne A
• Format: Journal Article
• Language: English
• Published: San Francisco Public Library of Science 30.08.2022; Public Library of Science (PLoS)
• Subjects: Adsorption; Amino acids; Antibodies; Antigens; Bioavailability; Biology and Life Sciences; Cell surface; Colonies; Colony-forming cells; Conjugation; Endothelial cells; Endothelium; Grafting; Health aspects; Immobilization; Immunoglobulin G; Implants; Medicine and Health Sciences; Physical Sciences; Polypeptides; Polystyrene; Polystyrene resins; Progenitor cells; Properties; Prostheses; Prosthetics; Protein G; Proteins; Research and Analysis Methods; Stem cells; Stent (Surgery); Stents; Substrates; Surface antigens; Surface chemistry; Surgical implants; Thrombosis; Transplants & implants; Vascular implants; Viral antibodies; Canada; United States > US; Quebec Canada
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0269316

Maximizing the re-endothelialization of vascular implants such as prostheses or stents has the potential to significantly improve their long-term performance. Endothelial progenitor cell capture stents with surface-immobilized antibodies show significantly improved endothelialization in the clinic. However, most current antibody-based stent surface modification strategies rely on antibody adsorption or direct conjugation via amino or carboxyl groups which leads to poor control over antibody surface concentration and/or molecular orientation, and ultimately bioavailability for cell capture. Here, we assess the utility of a bioaffinity-based surface modification strategy to immobilize antibodies targeting endothelial cell surface antigens. A cysteine-tagged truncated protein G polypeptide containing three Fc-binding domains was conjugated onto aminated polystyrene substrates via a bi-functional linking arm, followed by antibody immobilization. Different IgG antibodies were successfully immobilized on the protein G-modified surfaces. Covalent grafting of the protein G polypeptide was more effective than surface adsorption in immobilizing antibodies at high density based on fluorophore-labeled secondary antibody detection, as well as endothelial colony-forming cell capture through anti-CD144 antibodies. This work presents a potential avenue for enhancing the performance of cell capture strategies by using covalent grafting of protein G polypeptides to immobilize IgG antibodies.