Encapsulating Nerve Growth Factor in High-Density Lipoprotein-Mimicking Nanoparticles

Samenvatting

Source: Zhu, J., et. al., Preparation and Characterization of Novel HDL-mimicking Nanoparticles for Nerve Growth Factor Encapsulation. J. Vis. Exp. (2017)

This video demonstrates the stepwise assembly of NGF-loaded HDL-mimicking nanoparticles using protamine-assisted encapsulation and lipid-based self-assembly for potential neurotrophic delivery applications.

Protocol

1. Preparation of NGF HDL-mimicking Nanoparticles

1. Dissolve the excipients, phosphatidylcholine (PC), sphingomyelin (SM), phosphatidylserine (PS), cholesteryl oleate (CO), and D-α-tocopheryl polyethylene glycol succinate (TPGS), in ethanol to prepare stock solutions at 1 mg/mL.
   NOTE: The stock solutions were aliquoted and stored at -20 °C. The cholesteryl oleate was stored in dark bottles. The PC, SM, PS, and CO stock solutions were stable for up to 6, 3, 12, and 12 months, respectively, at -20 °C. The TPGS solution was stable for at least 12 months at -20 °C.
2. Mix 10 µL of NGF (1 mg/mL in water) with 10 ....

Materialen

Lijst van materialen gebruikt in dit artikel

Naam	Bedrijf	Catalogusnummer	Opmerkingen
Recombinant Human Beta-NGF	Creative Biomart	NGF-05H
L-a-Phosphatidylcholine (PC)	Avanti	131601P	95%, Egg, Chicken
Sphingomyelin (SM)	Avanti	860062P	Brain, Porcine
Phosphatidylserine (PS)	Avanti	840032P	Brain, Porcine
Cholesteryl oleate (CO)	Sigma	C9253
D-α-Tocopheryl polyethylene glycol succinate (TPGS)	BASF	9002-96-4	Vitamin E Polyethylene Glycol Succinate
Protamine sulfate	Sigma	P3369	meets USP testing specifications
Apolipoprotein A1, Human plasma	Athens Research & Technology	16-16-120101	1mg in 671 µl 10 mM ammonium bicarbonate, pH 7.4
Homogenizer	Tekmar	T 25-S1
Polytron homogenizer	Kinematica	PT 1200C