Methodological problems of direct bioluminescent ATP assay in platelets and erythrocytes

✍ Scribed by Girotti, Stefano ;Ferri, Elida ;Cascione, Maria Loredana ;Comuzio, Sergio ;Mazzuca, Augusto ;Orlandini, Alessandro ;Breccia, Alberto

• Publisher: John Wiley and Sons
• Year: 1989
• Weight: 548 KB
• Volume: 4
• Category: Article
• ISSN: 0884-3996
• DOI: 10.1002/bio.1170040178

✦ Synopsis

Direct bioluminescent ATP determination in platelets and erythrocytes involves the study of different parameters which are discussed here. Some parameters are linked to the bioluminescent reaction and t o the analyte (ATP); others have regard t o the biological matrix. The composition of bioluminescent reagents and the preparation and conservation of the ATP standard, also in the presence of excipients, are among the first given. Matrix problems involve cell characteristics related t o age and form, lysis resistance and the possible formation of aggregates (platelets) that may inhibit the complete release of ATP. For these reasons we used the most efficient ATP release agent with the lowest inhibitory effect on luciferase. The data obtained correlate well with a bioluminescent method requiring extraction with ethanol/EDTA, and therefore more time, for ATP determination in platelets and erythrocytes.