Identification of vitronectin as a major plasma protein adsorbed on polymer surfaces of different copolymer composition.
The arrays of proteins adsorbed from plasma onto a series of polystyrene copolymeric latexes were analyzed by enzyme-linked immunosorbent assay (ELISA) of washed beads and immunoblotting of proteins desorbed from the beads and separated by polyacrylamide gel electrophoresis (PAGE). Beads were prepared by continuous emulsion polymerization in the absence of surfactant. Coomassie brilliant blue staining of gel electropherograms of desorbed proteins indicated that the presence of small amounts of comonomers (1 to 10 mole %) significantly influenced the composition of the adsorbed protein layer. Immunoblotting revealed that fibrinogen, fibronectin, and vitronectin were adsorbed by all surfaces investigated. C3 and Clq adsorption varied significantly with copolymer composition. The ELISAs revealed that although the concentrations of vitronectin and fibronectin in plasma are similar, the extent of vitronectin adsorption from 70% to 85% plasma was greater by two orders of magnitude than fibronectin adsorption. Vitronectin adsorbed on carboxylic acid-containing copolymers reacted more strongly with a conformationally sensitive antivitronectin monoclonal antibody (MoAb) than vitronectin adsorbed to polystyrene and was more susceptible to cleavage by plasma proteases(s). The results show that vitronectin is a major protein adsorbed from concentrated plasma and that small changes in the chemical composition of a copolymer profoundly affects the extent and nature of protein adsorption from complex mixtures such as plasma.
Bale
M D
MD
Life Sciences Research Laboratories, Eastman Kodak Company, Rochester, NY 14650-2113.
Wohlfahrt
L A
LA
Mosher
D F
DF
Tomasini
B
B
Sutton
R C
RC
D000179:Acrylates; D000327:Adsorption; D015153:Blotting, Western; D015922:Complement C1q; D003176:Complement C3; D004797:Enzyme-Linked Immunosorbent Assay; D005340:Fibrinogen; D005353:Fibronectins; D006023:Glycoproteins; D006801:Humans; D066298:In Vitro Techniques; D008970:Molecular Weight; D011137:Polystyrenes; D013329:Structure-Activity Relationship; D019096:Vitronectin
D000179:Acrylates; D003176:Complement C3; D005353:Fibronectins; D006023:Glycoproteins; D011137:Polystyrenes; D019096:Vitronectin; D015922:Complement C1q; D005340:Fibrinogen