Surface Presentation of Bioactive Ligands in a Nonadhesive Background Using DOPA-Tethered Biotinylated Poly(ethylene glycol)

Abstract

We have developed surfaces for the selective presentation of biotinylated peptides and proteins in a background that resists nonspecific protein adsorption; controlled amounts of biotinylated polyethylene glycol (MW 3400; PEG3400) anchored to titanium-dioxide-coated surfaces via an adhesive tri-peptide sequence of L-3,4-dihydroxyphenylalanine (DOPA 3 -PEG3400-biotin; DPB) were incorporated within a DOPA 3 -PEG2000 background. Using optical waveguide lightmode spectroscopy, we found that the amounts of sequentially adsorbed NeutrAvidin and singlybiotinylated molecules increased proportionally with the amount of DPB in the surface. Biotinylated peptides (MW ca. 2000) were able to fill all three of the remaining avidin-binding sites, while only one molecule of biotinylated PEG5000 or stem cell factor bound to each avidin. The resulting biotinavidin-biotin linkages were stable for prolonged periods under continuous perfusion, even in the presence of excess free biotin. Hematopoietic M07e cells bound to immobilized peptide ligands for α5β1 (cyclic RGD) and α4β1 (cylic LDV) integrins in a DPB-dose-dependent manner, with nearmaximal binding to cylic LDV for surfaces containing 1% DPB. Multiple ligands were adsorbed in a controlled manner by incubating NeutrAvidin with the respective ligands in the desired molar ratio and then adding the resulting complexes to DPB-containing surfaces. Cell adhesion to surfaces containing both cylic LDV and cyclic RGD increased in an additive manner compared to that for the individual ligands. The bioactivity of adsorbed biotinylated stem cell factor was retained, as demonstrated by DPB-dose-dependent M07e cell adhesion and ERK1/2 activation.

Multifunctional biomaterial coatings: synthetic challenges and biological activity

“…Furthermore, streptavidin can bind several biotinylated moieties, creating a highly versatile system. Thus, this method is especially suitable for analytical investigations as shown in Figure 5F (Gunawan et al, 2007;Migliorini et al, 2014). Another affinity-based approach is the already described strategy that employs the self-organization of nucleic acids.…”

“…A non-covalent, affinity-based system was used to immobilize cRGD and cLDV on DOPA-modified PEG (Table 3, line h). Since avidin is able to bind several biotinylated peptides, both integrin-ligands could be displayed in a specific ratio, yielding a beneficial synergistic cell response (Gunawan et al, 2007). In addition to peptides, PEG functionalized surfaces were coated with proteins such as the anti-CD34 antibody to recruit endothelial progenitor cells from the blood stream (Table 3, line i).…”

Mussel-mimetic strong adhesive resin from bio-base polycoumarates

“…However, potential applications of this adhesive function have fascinated many researchers. For example, the group of Messersmith and Miller [8][9][10][11][12] has demonstrated to mimic a material that mimics the behavior of mussel adhesive in the form of adhesive sheets. Although this strong adhesion has been attributed to hydrogen bond interactions or covalent bonding between the catechol groups, the adhesive mechanism is not completely understood.…”

Effects of adhesive characteristics of the catechol group on fiber-reinforced plastics

“…6,7 However, these functions have fascinated many researchers regarding applications. For example, the research group of Messersmith and Miller, [8][9][10][11][12] has mimicked the adhesion for application in adhesive sheets, and we have successfully synthesized mussel-mimetic high-performance adhesive resins. 13 This finding indicates that the molecular interactions between catechols and inorganic surfaces can be used to develop macroscopic materials and has motivated us to utilize this catechol interaction to develop organic-inorganic hybrids such as glass-fiberreinforced plastics.…”