Polypeptide films and methods
A polypeptide and multi-layer film technology, applied in the direction of microcapsules, coatings, microcapsule preparations, etc., can solve problems such as attacks
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Embodiment 1
[0077] Example 1: Adsorption of glucose oxidase (GOx) to CaCO 3 and MF particles on
[0078] Considering the use of the enzymatic properties of GOx, GOx was chosen as the template protein. In the GOx adsorption experiment, 5 mg CaCO 3 Particles (PlasmChem GmbH, Germany) or melamine formaldehyde (MF) particles were mixed with 100 μL of 10 mM Tris buffer (pH 7.4) containing 37,300 U / g Type II-S GOx from A. niger (SIGMA, USA). The temperature for GOx adsorption is the temperature at which the GOx solution remains in a liquid state (for pure water, at a pressure of about 1 atm, the nominal temperature is about 0-100°C), and at this temperature, the enzymatic activity of GOx will not be irreversibly inactivated . The final particle concentration was 5% (w / v). Use Jasco V-430 spectrophotometer (Japan), 280nm, measure the liquid phase absorbance, and determine the adsorption amount of the enzyme on the particle surface by the decrease of the absorbance. Centrifuge to separate th...
Embodiment 2
[0079] Example 2: Encapsulation of enzymes into polypeptide microcapsules
[0080] In the encapsulation experiment study of this example, since it is easy to buy poly(L-lysine) (PLL) (MW about 14.6kDa) and poly(L-glutamic acid) (PLGA) (MW about 13.6kDa) from the market , so they were selected as oppositely charged polypeptides for layer deposition operations. The process of encapsulating with designed peptides instead of PLL and / or PLGA is basically similar. An example of a pair of oppositely charged peptides that exert the same effect is described in Li and Haynie (2004) Biomacromolecules 5: 1667-1670.
[0081] Adsorption of GOx to CaCO 3 The operation on the microparticles is as follows: to 5mg CaCO 3 Add 0.10 mL of 5 mg / mL GOx solution to the microparticles, mix well for 2 hours, centrifuge at 1000×g for 5 minutes, and remove the liquid phase.
[0082] Multilayer deposition of each peptide onto GOx-adsorbed CaCO 3 The manipulation process on the microparticles included...
Embodiment 3
[0090] Example 3: Encapsulation of enzymes into polyelectrolyte microcapsules
[0091] In another embodiment, hemoglobin was loaded onto calcium carbonate particles in phosphate buffered saline (pH 7.0), and then encapsulated with non-peptide polyelectrolytes. Adding a polymer precipitant such as 40% PEG300 to the protein solution increases the protein loading efficiency. The adsorbed protein was encapsulated by the LBL process with two oppositely charged polyelectrolytes. Suitable polymers for this example are poly(styrene sulfonate), polyanion and poly(allylamine), polycation. The loading of the protein can be demonstrated with a spectrophotometer in the visible range: since hemoglobin contains heme, there is a large absorption spectrum near 410 nm.
[0092] The invention proposes a method for efficiently retaining functional biologically active molecules in nano-engineering polyelectrolyte membranes by adding a polymer precipitant to the assembly buffer. Polyelectrolyte ...
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