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Thrombin binding properties of insoluble modified polystyrene: Part II
A M Fischer1, J Tapon-Bretaudiere, A Bros
1Département d'Hématologie, C.H.U. Necker-Enfants Malades, Paris, France.
Thrombosis Research
|April 15, 1988
Summary
New affinity chromatography resins effectively purify thrombin. These arginyl-grafted polystyrene resins show high specificity and retain thrombin
Area of Science:
- Biochemistry
- Protein Chemistry
- Affinity Chromatography
Background:
- Antithrombin III (AT III) regulates thrombin activity through arginine-serine interactions.
- Developing specific methods for thrombin purification is crucial for biochemical research and therapeutic applications.
Purpose of the Study:
- To synthesize and characterize novel insoluble polystyrene resins grafted with arginyl methyl ester for thrombin affinity chromatography.
- To evaluate the specificity and efficiency of these resins in binding and purifying human thrombin.
Main Methods:
- Synthesis of arginyl methyl ester-grafted polystyrene resins.
- Affinity chromatography experiments comparing thrombin binding with other proteins like prothrombin, Factor IXa, trypsin, and AT III.
- Elution studies using varying ionic strengths and analysis of thrombin activity post-purification.
Main Results:
- A selected resin demonstrated high affinity for thrombin, binding 0.7 mg per gram of resin.
- Elution required high ionic strength (1.5 M), with preserved amidolytic and clotting activities.
- The resin exhibited specificity for thrombin, showing minimal binding to other tested serine proteases and precursors.
Conclusions:
- Arginyl-grafted polystyrene resins provide an efficient and specific method for thrombin purification via affinity chromatography.
- This technique is valuable for isolating thrombin and removing it as a contaminant from plasma protein fractions.