Encapsulation of biologically active agents
A technology of substance and drug, applied in the field of encapsulation of bioactive agents
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[0049] The present invention provides a particulate carrier comprising a particle-forming substance and a bioactive agent, and a method for preparing the particulate carrier.
[0050] In one embodiment, the nanoparticles of the invention comprise bioactive agents such as proteins or peptides. The protein may be an antigen binding molecule, which as used herein refers to antibodies, antibody fragments and other protein structures capable of binding a target.
[0051] Antigen binding molecules may include domains. A "domain" is a folded protein structure that has a tertiary structure independent of the rest of the protein.
[0052] In general, domains are responsible for discrete functional properties of proteins, and in many cases can be added, removed or transferred to other proteins without loss of function of the protein and / or the remainder of the domain. A "single antibody variable domain" is a folded polypeptide domain that comprises the sequence characteristics of an a...
Embodiment 1
[0186] The polymerization of embodiment 1BCA (butyl cyanoacrylate) monomer
[0187] Polymers are formed by rapid polymerization in organic solvents:
[0188] BCA monomer (200 μl, Vetbond, 3M) was added to 1 ml absolute ethanol in a 25 ml beaker and the beaker was swirled slowly. The resulting solution was mixed gently until polymerization started. A white solid dispersion was formed after the polymerization was complete. Mixing of the dispersion was stopped when the reaction mixture became too viscous to stir.
[0189] The ethanol in the reaction mixture was allowed to evaporate for at least 1 h in a fume hood. After evaporation of ethanol, a cracked white solid mass was obtained. This solid was collected and used in the nanoparticle preparation process.
Embodiment 2
[0190] Embodiment 2 prepares hollow nanoparticles by double emulsion method
[0191] The PBCA polymer was dissolved in ethylene dichloride at a concentration of 1% w / v, and was used to prepare hollow PBCA by emulsifying into a double emulsion (water / oil / water, w / o / w) as follows Nanoparticles:
[0192] (i) Primary emulsification (w / o)
[0193] Internal phase (w): 5% sodium cholate (SIGMA) in water or buffer prepared by the following mixing process:
[0194] 500 μl of water or buffer; and
[0195] 500 μl sodium cholate (10% w / v stock solution).
[0196] The total volume of the internal aqueous phase was 1 ml. Keep the solution on ice until ready to use. Before use, each solution was drawn into an insulin syringe (Terumo 1 ml, BDmicrolanceneedle 19G1.5").
[0197] External (organic) phase (o): PBCA polymer (1% w / v) in dichloromethane (DCM, Fischer).
[0198] The organic phase (PBCA polymer in DCM, 6ml) was poured into a 10ml beaker (cooled on ice) and the probe of the homo...
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Abstract
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