Antibody binding, platelet adhesion, and protein adsorption on various polymer surfaces

Abstract

Commercially available polymer surfaces for in vitro applications are characterized to different extents in terms of hydrophobicity, binding preferences, and immunoglobulin capacity. We compared five, well or poorly defined polystyrene plates, used as standard hydrophobic surfaces for studying biological interactions. Antibody binding (ELISA) and platelet adhesion (release of alkaline phosphatase from adhered platelets) were contrasted with total protein adsorption (alkaline phosphatase assay and bicinchoninic acid assay). In the assays, we utilized four plasma proteins: human serum albumin (HSA), C-reactive protein (CRP), fibronectin, and fibrinogen. At 0.5 μg antigen/well, all antibodies bound to their antigens most effectively on Nunc (MaxiSorp and MediSorp, Waltham, Massachusetts, USA) microplates, as compared to Sarstedt (Nümbrecht, Germany) and Corning microplates (Tewksbury, Massachusetts, USA). The significant differences between Nunc and Corning were seen in the binding of anti-HSA (P ≤ 0.01) and anti-fibronectin (P ≤ 0.0002). Similar patterns were shown in experiments of ADP-induced platelet adhesion to fibrinogen immobilized at 200 μg/well. Platelet adhesion noted on Corning microplates was roughly three times lower compared to those observed on MaxiSorp (P < 0.01), MediSorp (P < 0.02), and Sarstedt (P < 0.05). In a parallel study, we have also shown the superiority of tissue culture-Sarstedt surface over micro test Sarstedt plate. Furthermore, the antibody binding, but not platelet adhesion, was positively correlated with total protein adsorption. Our findings indicate that of five polystyrene surfaces, Nunc microplates are optimal for studies of protein adsorption, as they had the highest binding capacity and relatively the least affected protein structure, pointing to the role of surface chemistry in protein adsorption and adsorption-induced conformational changes in a protein structure.

Protein Interactions with Polymer Coatings and Biomaterials

“…In addition to using diluted protein solutions for mechanistic studies, it is particularly interesting to apply application‐relevant concentrations, e.g., physiological concentrations, for predictive studies of competition and exchange. For example, at cell culture plates with different proposed binding capabilities, fibronectin only showed differences in adsorption at low protein concentrations, but these differences were insignificant at higher concentrations 111. While the supernatant of polymer samples with the nonadsorbed protein fraction is easily accessible, higher precision may be assigned to a characterization of the adsorbed protein fraction.…”

“…by gel chromatography), their direct selective biochemical detection (e.g. with antibodies,111 aptamers124, 125), or indirect detection (after labeling) 126–128…”

An asymmetrical dual coating on the stent prepared by ultrasonic atomization

“…So that the ADC sprayed on the stent can solve a series of side effects of hemolysis, coagulation and inflammation response which caused by plasma protein and platelet adhesion and aggregation on the stent after stent implantation 28 . We also found that the ADC could significantly prolong the APTT, PT and TT and reduce the amount of Fbg adsorption compare with other groups, which indicating that SZ-21 can slow down the clotting system of endogenous and exogenous system, and some studies have shown that Fbg adhesion reduction also inhibits thrombosis [29][30][31] , so the ADC have good blood compatibility. Moreover, the hemolysis rate of ADC was less than 5 % which meet the standards that clinical use of biological material requires 32 .…”

Wechselwirkungen von Proteinen mit Polymerbeschichtungen und Biomaterialien

“…Es konnte z. B. an Zellkulturplatten mit verschiedenen bereitgestellten Bindungskapazitäten gezeigt werden, dass Fibronektin nur bei geringer Konzentration Unterschiede im Adsorptionsverhalten aufweist und dass diese Unterschiede bei höherer Konzentration unbedeutend sind 111. Während der Überstand einer Polymerprobe mit dem Anteil der nicht‐adsorbierten Proteine leicht zugänglich ist, ist eine höhere Genauigkeit bei der Charakterisierung des adsorbierten Polymerteils notwendig.…”

“…B. durch Gelchromatographie, oder ihren direkten selektiven biochemischen Nachweis, z. B. mithilfe von Antikörpern111 oder Aptameren,124, 125 oder indirekten Nachweis (nach einer Markierung) 126–128…”