Kits, formulations and solutions having enzymatically-permissive amounts of visualization agents and uses thereof
一种显像剂、试剂盒的技术，应用在蛋白水解酶组合物领域，能够解决没有公开染料特定浓度等问题
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0128] Example 1: Effect of different dyes on thrombin coagulation activity.
[0129] The purpose of this study was to determine the effect of dyes added to fibrin glue formulations on thrombin activity. For this purpose, thrombin similar to the two-component fibrin sealants described in US-B-6,121,232 and WO9833533 was formulated using different dyes at a final concentration of 0.01-0.2%. The compatibility of the dye with thrombin was determined by measuring the coagulation activity of thrombin in different preparations according to the modified European Pharmacopoeia method (0903 / 1997) described below.
[0130] Briefly, standard solutions of thrombin (4, 6, 8 and 10 IU / ml) or test samples were incubated at 30°C for 2 minutes. Then 40 μl of thrombin solution of each solution was mixed with 160 μl of fibrinogen solution (0.1%; Enzyme research; cat No FIB1 2800L) and the clotting time was determined. Use the standard to make a calibration curve with Log clotting time versus l...
Embodiment 2
[0136] Example 2: Effect of different dyes on clotting kinetics.
[0137] Human thrombin was mixed with different dyes to a final dye concentration of 0.005-0.2%. The effect of dyes on clotting kinetics was determined using a drip coagulation test model. Briefly, fibrin coagulation kinetics was measured using a 7 × 10 5 It is carried out on the slope of the device driven by Pa nitrogen pressure. In each experiment, 5-fold dilutions of 5ml bioactive component (BAC) and 5ml thrombin solution (final: 200IU / ml) (in 40mM CaCl 2 Middle) pumped into a separate syringe. BAC was prepared from the concentrated cryoprecipitate after establishment as disclosed in EP-A-534 178 (where arginine and tranexamic acid were added as described in US-B-6,121,232 and WO-A-9833533). The two solutions are released simultaneously (approximately 1 / 8 of each), and the mixed droplet is dripped on the slope, and the droplet seeps down the slope until a clot is formed. The distance traveled by the drople...
Embodiment 3
[0148] Example 3: Stability of coagulation activity of fibrin glue with imaging agent (dye) after freeze-thaw (F&T).
[0149] Human thrombin was formulated with methylene blue, crystal violet, bromothyme blue or riboflavin at a final concentration of 0.005-0.1%. The different formulations were flash frozen to -35°C and subsequently thawed. The effect of the freeze-thaw operation on the coagulation activity of thrombin was evaluated using the drip coagulation test model (migration time test as described in Example 2). The results are summarized in Table 3.
[0150] Table 3: Effect of freeze-thaw operation on gel stability
[0151]
[0152] *NA - not available
[0153] The coagulation activity of fibrin glue formulated with methylene blue did not change significantly after freeze-thaw operations. Thus, this experiment shows that the coagulation activity of fibrin glue formulations containing methylene blue is stable even through freeze-thaw.
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More