The effect of proteolytic degradation of plasma fibronectin on the responses of functional and immunometric assays for intact fibronectin.
Numerous studies have been made into the nature and importance of altered levels of plasma fibronectin seen in a range of clinical disorders. However, fibronectin is highly sensitive to the proteolytic degradation which may accompany some of these conditions. The influence of such degradation on the assays used is frequently unknown. We have investigated the effects of controlled protease degradation on the responses of an electroimmuno and a functional assay, using purified fibronectin. Tryptic digestion influenced the assays more than thrombin, in relation to the degree of breakdown. Both enzymes reduced the functional (gelatin-binding) activity whilst tryptic cleavage increased the apparent concentration by electroimmunoassay. The findings demonstrate that certain assays may be inappropriate for those clinical conditions where levels of intact fibronectin are of most interest. It is also necessary, then, to determine the degree of fibronectin breakdown when measuring pathological levels.
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005353:Fibronectins / Q000097:blood*; D005780:Gelatin / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D007122:Immunoelectrophoresis; D010447:Peptide Hydrolases / Q000378:metabolism*; D011485:Protein Binding; D013917:Thrombin / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
D005353:Fibronectins; D005780:Gelatin; D010447:Peptide Hydrolases; D014357:Trypsin; D013917:Thrombin