Effect of lecithin and MgCO3 as additives on the enzymatic activity of carbonic anhydrase encapsulated in poly(lactide-co-glycolide) (PLGA) microspheres.
A model enzyme, carbonic anhydrase, was encapsulated and released from poly(lactide-co-glycolide) (PLGA) microspheres (1-3 microm) made by a novel phase inversion technique. Lecithin was used as a surfactant in the encapsulation process and was incorporated in either the organic phase, aqueous phase, both phases, or not at all. Additional microspheres were also made with lecithin incorporated in the aqueous phase and a basic salt, MgCO3, in the polymeric phase. Released carbonic anhydrase, protein extracted from microspheres, or enzyme incubated with lecithin and PLGA were analyzed via HPLC and activity assay to determine the effect of these additives on protein integrity and activity. Lecithin in the aqueous phase appeared to increase the fraction of enzyme in monomeric form as well as its activity for both extracted protein and released protein as compared to the other formulations without MgCO3. Incubation of enzyme with PLGA degradation products indicated that the acidic environment within the microspheres aids in the irreversible inactivation of the encapsulated protein. Addition of MgCO3 further increased the amount of monomer in both the extracted and released protein by decreasing the amount of acid-induced cleavage and noncovalent aggregation, but still greatly decreased the activity of the enzyme.
Sandor
Maryellen
M
Department of Molecular Pharmacology, Physiology, and Biotechnology, Brown University, Providence, RI 02912, USA.
Riechel
Alex
A
Kaplan
Ian
I
Mathiowitz
Edith
E
D002256:Carbonic Anhydrases / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D016503:Drug Delivery Systems; D004789:Enzyme Activation; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D019344:Lactic Acid; D008274:Magnesium / Q000737:chemistry; D008855:Microscopy, Electron, Scanning; D008863:Microspheres; D010446:Peptide Fragments / Q000032:analysis; D010713:Phosphatidylcholines / Q000737:chemistry; D011100:Polyglycolic Acid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers; D012995:Solubility; D017550:Spectroscopy, Fourier Transform Infrared
D010446:Peptide Fragments; D010713:Phosphatidylcholines; D011108:Polymers; C005479:magnesium carbonate; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D002256:Carbonic Anhydrases; D008274:Magnesium