Method of quality control testing a Factor XIII containing sample
The invention relates to a method of quality control testing a Factor XIII (FXIII) containing sample which comprises the step of detecting the presence of and/or measuring the concentration of pre-activated FXIII (FXIIIao) in said sample and to a quality control kit for determining the quality of a Factor XIII (FXIII) containing sample. Preferably, an anion-exchange chromatographic column is used, as well as the fluorescent substrate Abz-NE (Cad-Dnp) EQVS PLTLLK-OH.
1. A method of quality control testing a sample containing Factor XIII (FXIII) by determining the level of pre-activated FXIII in the sample, comprising:
measuring activated FXIII in the sample in the absence of thrombin and in the presence of 0.5-10 mM atomic absorption standard Ca 2+ ; and
measuring activated FXIII in the sample in the presence of thrombin and 0.5-10 mM atomic absorption standard Ca 2+ ,
wherein the ratio of (i) activated FXIII in the absence of thrombin to (ii) activated FXIII in the presence of thrombin indicates the level of pre-activated FXIII in the sample.
2. The method according to claim 1 , wherein the measuring steps comprise measuring the activated FXIII in the presence of 1-2 mM atomic absorption standard Ca 2+ .
3. The method according to claim 2 , wherein the measuring steps comprise measuring the activated FXIII in the presence of 2 mM atomic absorption standard Ca 2+ .
4. The method according to claim 1 , wherein the measuring steps are performed in the presence of a fluorometric or non-fluorometric substrate.
5. The method according to claim 4 , wherein the substrate comprises Abz-NE(Cad-Dnp)EQVSPLTLLK.
6. The method according to claim 1 , wherein the measuring steps further comprise measuring the activated FXIII in the presence of 50-500 mM NaCl.
7. The method according to claim 6 , wherein the measuring steps further comprise measuring the activated FXIII in the presence of 100-200 mM NaCl.
8. The method according to claim 1 , wherein the measuring steps further comprise measuring the activated FXIII at a pH between 6.5 and 10.
9. The method according to claim 8 , wherein the measuring steps further comprise measuring the activated FXIII at a pH between 7 and 8.
10. The method according to claim 1 , wherein the measuring steps further comprise measuring the activated FXIII in the presence of 50-200 mM Hepes buffer.
11. The method according to claim 10 , wherein the measuring steps further comprise measuring the activated FXIII in the presence of 100 mM Hepes buffer.
12. The method according to claim 1 , wherein the measuring steps further comprise measuring the activated FXIII in the presence of a surfactant.
13. The method according to claim 12 , wherein the surfactant is a polyethylene glycol.