Description
PURPOSE
Drug contamination by pyrogens is a serious threat to patient health and can lead to fever and eventually shock and death. The Monocyte Activation Test (MAT) is a new method for the detection of the full range of pyrogenic substances, including bacterial endotoxins and pyrogens of non-endotoxin nature (peptidoglycans, yeasts, fungi, viruses). The test is a complete replacement for the rabbit pyrogen test and eliminates the use of animals.
«ALPYR MAT» Monocyte Activation Test kit is used for pyrogen testing in medicines, API and medical devices by quantitative, semi-quantitative assays and by comparative assay with a control series which is approved as a standard. The kit allows to detect endotoxins with high sensitivity and accuracy without using animals.
«ALPYR MAT» Monocyte Activation Test kit includes the following components:
| Component name | Quantity per set | Storage conditions |
| MAT cells (cryopreserved peripheral blood monocytes, 10 million cells/ml) | 1x1 ml | -80 °C |
| Cell culture supplement FBS (ultra-low endotoxin fetal bovine serum) | 1x3 ml | -20 °C |
| Cell culture medium RPMI (contains L-glutamine, sterile) | 1x50 ml | 2-8 °C |
| Endotoxin standard (E.Coli lipopolysaccharide, 2000 IU/ml) | 1x50 µl | -20 °C |
| Non-endotoxin control LTA (lipoteichoic acid, 2 mg/ml) | 1x50 µl | -20 °C |
| Non-endotoxin control HKSA (heat killed Staphylococcus aureus, 1010 HKSA/ml) | 1x20 µl | -20 °C |
| ELISA kit for detection of interleukin-6 (detection range 10-3160 pg/ml) | 1 pc | 2-8 °C |
| Sterile flat-bottom 96-well plate | 1 pc | 2-30 °C |
PRINCIPLE OF THE METHOD
The method is based on the ability of monocyte blood cells to produce the human inflammatory mediator interleukin-6 in the presence of pyrogenic substances. The assay is run in two steps. At the first step monocytes are incubated along with the test sample for about 20 hours. In the presence of pyrogens in the test sample, monocytes produce cytokines (including interleukin-6). In the second step, the amount of released interleukin-6 is determined using enzyme-linked immunosorbent assay (ELISA). A calibration curve is plotted using endotoxin standard solutions. The amount of interleukin-6 in the cell supernatant is directly proportional to the amount of pyrogens in the samples.
SAMPLES TO BE TESTED
- pharmaceuticals;
- active pharmaceutical ingredients;
- washes from medical devices;
- biotech products;
- cell therapy products;
- immunobiologicals – vaccines, serums, albumins.
FEATURES
- detection of the full range of pyrogens;
- high sensitivity of the kit – up to 0.015 EEU/ml;
- does not require the use of animals;
- international standard endotoxin (LPS of Gram-negative bacteria) and non-endotoxin controls (lipoteichoic acid and heat killed Staphylococcus aureus) are included in the kit;
- storage temperature of the vial with monocytes -80 °C and below.
