Glass-bead affinity chromatography of cell attachment and spreading-promoting factors of human serum.
The glass-binding properties of a number of purified glycoproteins capable of promoting attachment and spreading of a variety of types of animal cells in culture have been examined. Two such factors in human serum, fibronectin and serum spreading factor, exhibited strong affinities for glass beads and could be eluted from glass-bead columns under similar conditions. A number of other glycoproteins of human serum that do not promote cell adhesion did not bind to glass beads under conditions that resulted in binding of serum spreading factor or fibronectin. At a sufficiently low ratio of serum volume to glass-bead volume, human serum could be simultaneously depleted of serum spreading factor, fibronectin, and cell spreading-promoting activity by glass-bead affinity chromatography. Laminin, another cell spreading-promoting glycoprotein, possessed glass-binding properties similar to those of serum spreading factor and fibronectin while chondronectin, a fourth cell spreading-promoting factor of more limited specificity of biological activity and distribution in vivo, did not exhibit a strong interaction with glass beads under the same conditions. These observations suggest that glass-bead column affinity chromatography may prove useful as a general method for isolation and study of glycoprotein factors promoting attachment and spreading of cells in culture.
D000818:Animals; D001798:Blood Proteins / Q000032:analysis*; D002448:Cell Adhesion; D002846:Chromatography, Affinity / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D005353:Fibronectins / Q000302:isolation & purification*; D005898:Glass; D006023:Glycoproteins / Q000302:isolation & purification*; D006367:HeLa Cells / Q000032:analysis; D006801:Humans; D007797:Laminin / Q000302:isolation & purification; D011506:Proteins / Q000302:isolation & purification; D012513:Sarcoma, Experimental / Q000032:analysis; D013056:Spectrophotometry, Ultraviolet; D014020:Tissue Extracts / Q000032:analysis; D019096:Vitronectin; D005719:gamma-Globulins / Q000032:analysis
D001798:Blood Proteins; D005353:Fibronectins; D006023:Glycoproteins; D007797:Laminin; D011506:Proteins; D014020:Tissue Extracts; D019096:Vitronectin; C029172:chondronectin protein, human; D005719:gamma-Globulins