Determination of protease activity in blood and microorganisms

Autor: A. A. Mishin, V. L. D’yakov, N. I. Leont’eva, L. I. Novikova, L. V. Kozlov, A. M. Bichucher, N. M. Gracheva
Rok vydání: 2008
Předmět:
Zdroj: Biochemistry (Moscow) Supplement Series B: Biomedical Chemistry. 2:381-384
ISSN: 1990-7516
1990-7508
DOI: 10.1134/s1990750808040082
Popis: A protease activity may be determined by means of immunoglobulins. Since proteolytic products apparently do not retain antigenic determinants of the initial substrate, the monitoring of enzymatic process may employ ELISA methods. The ELISA determination of functional activity of specific IgA1 protease has been used not only for detection of this enzyme, but also for measurement of its inhibition constants. IgG adsorbed onto a microplate was used for evaluation of total proteolytic activity. Varying pH values of the reaction medium it is possible to measure activity of neutral, alkaline and acid proteases. This approach was used for estimation total proteolytic activity of neutral proteases in blood serum. Due to high sensitivity of this method it was possible to dilute serum up to the level when serum inhibitors had not blocked enzyme activity. Assay of serum enzyme activity at acidic pH results in activation of pepsinogens and determination of pepsin activity. Measurement of a total level of serum pepsinogen activity may have diagnostic importance in gastroenterology, due to decisive contribution of pepsinogen I to the detectable activity.
Databáze: OpenAIRE
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