Protein-loaded PLGA microparticles engineered by flow focusing: physicochemical characterization and protein detection by reversed-phase HPLC.
In the present study, a novel synthesis technique based on the flow focusing (FF) technology is investigated for the preparation of green fluorescent protein (GFP)-loaded poly(D,L-lactide-co-glycolide) (PLGA) microparticles. To our knowledge, this novel technique has never been applied to the formulation of proteins in polymeric systems. A simple, specific and rapid reversed-phase HPLC (RP-HPLC) method was validated for the determination of GFP in PLGA microparticles with the best chromatographic peak resolution, reduced run time and low cost of analysis. In order to achieve the finest GFP-loaded polymeric particles, experimental parameters mainly associated to the FF device were studied (liquid flow rate and pressure of the focusing air). Very high GFP encapsulation values (>90%) were obtained by this technique, and the electrokinetic characterization of these systems suggested that this protein was incorporated into the polymeric matrix. This study is intended to offer information on which to base the development of high molecular weight protein-loaded polymeric delivery systems prepared by FF.
Holgado
M A
MA
Department of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of Seville, Spain. holgado@us.es
Cózar-Bernal
M J
MJ
Salas
S
S
Arias
J L
JL
Alvarez-Fuentes
J
J
Fernández-Arévalo
M
M
D001672:Biocompatible Materials / Q000737:chemistry; D055598:Chemical Phenomena; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004337:Drug Carriers / Q000138:chemical synthesis / Q000737:chemistry; D004339:Drug Compounding / Q000295:instrumentation* / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000737:chemistry*; D019344:Lactic Acid / Q000737:chemistry; D011100:Polyglycolic Acid / Q000737:chemistry; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer
D001672:Biocompatible Materials; D004337:Drug Carriers; D049452:Green Fluorescent Proteins; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid