Abstract
A reliable method for the measurement of different plasminogen activators is of great interest for both manufacturing and clinical medicine. A one-step assay based on a thickness shear mode acoustic sensor has been developed for this purpose. Two separate mixtures of substrates (fibrinogen and plasminogen) and enzymes (thrombin and the plasminogen activator) were mixed, and placed on the acoustic sensor surface. During the assay, the resonant frequency of a quartz crystal oscillating in the thickness shear mode was measured and used to find a characteristic clot dissolution time, from the sample addition to the time at the maximum dissolution rate. Calibrations of the acoustic assay were done for tissue-type plasminogen activator (t-PA) as well as for the other plasminogen activators: urokinase (u-PA); streptokinase (SK) and staphylokinase (SAK). All gave relative standard deviations of about 12%. Since the same method was used for all of the activators, their activities were compared, resolving the differences between their unit definitions. Linear relationships were found between urokinase and streptokinase which activate plasminogen directly and between t-PA and staphylokinase which require fibrin as a cofactor. The relationship between the groups was found to curve, indicating the difference between the two mechanisms. The acoustic method, therefore, may be used as a rapid and cost-effective reference method for the standardization and comparison of different plasminogen activators.
Similar content being viewed by others
References
Rouf SA, Moo-Young M, Chisti Y (1996) Biotechnol Adv 14(3):239–266
Rescigno AM, Waisman DM (2003) Plasminogen: structure, activation and regulation. Kluwer, New York
Collen D (1996) Circulation 93(5):857–865
Verheijen JH (1988) Purification, assay, and standardization of t-PA. In: Kluft C (ed) Tissue-type plasminogen activator (t-PA): physiological and clinical aspects. CRC Press, Boca Raton
Gronow M, Bliem R (1983) Trends Biotech 1:26–29
Marsh NA, Gaffney PJ (1977) Thromb Haemost 38(2):545–551
Johnson AJ, Kline DL, Alkjaersig N (1969) Thromb Diath Haemorrh 21(2):259–272
Saint-Denis BT, Humbert G, Gaillard JL (2001) J Dairy Res 68(3):437
Longstaff C, Whitton CM (2004) J Thromb Haemost 2(8):1416–1421
Gram J, Jespersen J, Declerck P, Kluft C (1999) Fibrinolysis Proteolysis 13:30–35
Sands D, Whitton CM, Longstaff C (2004) J Thromb Haemost 2(8):1411–1415
Sands D, Whitton CM, Merton RE, Longstaff C (2002) Thromb Haemost 88(2):294–297
Hayward GL, Dutton RL, Zhang Z, Scharer JM, Young MM (1998) Anal Commun 35:25–27
Longstaff C, Whitton C, Thelwell D (2007) J Thromb Haem 5(2):412–414
Lucklum R, Hauptmann P (2006) Anal Bioanal Chem 384(3):667–682
Ghazali M, Hayward GL (2008) The Analyst 133:910–913
Yao S, Mo Z (1987) Anal Chim Acta 193:97–105
Hayward G (1992) Anal Chim Acta 264(1):22–30
Czanderna AW, Lu C (1984) Applications of piezoelectric quartz crystal microbalances. Elsevier, Amsterdam
Kanazawa KK, Gordon JG (1985) Anal Chim Acta 175:99–105
Acknowledgement
We wish to thank the Natural Science and Engineering Research Council of Canada (NSERC) CellNet research network and discovery grant programs for financially supporting this research.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Ghazali, M., Hayward, G.L. Acoustic determination of performance and equivalence of plasminogen activators. Anal Bioanal Chem 392, 897–902 (2008). https://doi.org/10.1007/s00216-008-2343-y
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00216-008-2343-y