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AssigneeMerck Patent GmbH
InventorsUwe Wurzburg, Norbert Hennrich, Hans-Dieter Orth, Hermann Lang
Estimating the term…
A process for determination of CK-MB activity in a sample of a body fluid comprising the steps of incubating the sample, optionally in the presence of CK substrates, with antibodies which inhibit enzymatic activity of the M subunit in CK isoenzymes MM and MB without inactivating enzymatic activity of the B subunit in any CK-MB which may be present and determining the activity of the CK B subunit in the resultant sample.
A method for determining the enzymatic activity of creatinekinase-MB in a biological liquid sample, which comprises the steps of:
A composition for determining the activity of creatinekinase-MB in a liquid biological sample comprising antibodies being substantially free of CK-BB isoenzyme activity and being able to inhibit completely the enzymatic activity of the M subunit of creatinekinases MM and MB in solution without precipitation and without inactivating the enzymatic activity of the B subunit of any creatinekinase-MB in the sample, wherein complete inhibition of said M subunit leaves less than 5 U./l. of said enzymatic activity intact and non-inhibition of said enzymatic activity of said B subunit is less than 10 U./l., and an enzyme-coenzyme and substrate reagent for determining CK activity.
A composition for determining the activity of creatinekinase-MB in a liquid biological sample comprising
A test kit for the determination of the activity of creatinekinase-MB in a biological sample, which comprises:
A test kit for the determination of the activity of creatinekinase-MB in a biological sample, which comprises:
Dependencies are inferred from the claim text.
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Involving transferase: involving creatine phosphokinase
Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing:…
Chemistry: molecular biology and microbiology: packaged device or kit
Chemistry: molecular biology and microbiology: prevention or removal of interfering materials or reactants or other treatment to enhance results,…
Term ended 10 Jan 1995 (estimated)
USPTO file history isn’t available for this patent, so only indexed dates are shown.
The invention relates to a process and a composition for determining the activity of creatinekinase-MB in human body fluids.
The determination of the activity of creatinekinase (ATP: creatine-phosphotransferase, E.C. 2.7.3.2; abbreviation; CK) in serum is considered the most sensitive laboratory method for diagnosing diseases of skeletal muscles and the myocardium, especially myocardial infarction. However, differentiation between trauma of skeletal muscles and the myocardium is difficult, especially in making a differential diagnosis of myocardial infarction. Determination of total CK activity results in unreliable differentiation.
Prior attempts to improve the reliability of evidence provided by determination of CK activity in differential diagnosis have used measurements of activity of other enzymes in the serum and correlation of the resulting measurement with each other, for example, quotient CK/glutamic oxalacetic transaminase. However quotients of this type cannot be used to differentiate between cardiac infarction and pulmonary infarction or between cardiac infarction and secondary shock resulting from other causes.
Pure CK-MM, CK-MB or CK-BB is added to human serum inactivated in respect of its own CK activity and CK activities of individual samples are determined. 0.1 ml. of anti-CK-MM solution, obtained according to Example Ba, is then added to 0.1 ml. of a sample and the two are mixed and incubated for 5 minutes at 25.degree. C. Residual CK activity is then determined in a known way. Results are given in the table:
TABLE ______________________________________ Residual activities of CK isoenzymes after incu- bation with inhibiting anti-human-CK-MM (average values .+-. l s obtained from 5 determinations in each case) (s= standard deviation) Residual activity Activity of the after incubation isoenzymes added with anti-CK-MM Isoenzyme (U./l.) (U./l.) ______________________________________ CK-MM 98 .+-. 1.9 0.3 .+-. 2.1 1043 .+-. 22 0.5 .+-. 2.5 CK-MB 103 .+-. 2.0 53 .+-. 1.7 410 .+-. 7.8 206 .+-. 6.2 CK-BB 197 .+-. 3.8 199 .+-. 4.1 ______________________________________
Within errors of measurement, inhibition of activity is 100% for CK-MM, 0% for CK-BB and 50% for CK-MB, corresponding to the content of 50% of M subunits. Results are constant over a wide range of activities of isoenzymes added.