title: Stabilization of a model formalinized protein antigen encapsulated in poly(lactide-co-glycolide)-based microspheres.
abstract: A formaldehyde-mediated aggregation pathway (FMAP) has been shown to be primarily responsible for the solid-state aggregation of lyophilized formalinized protein antigens [e.g., tetanus toxoid (TT) and formalinized bovine serum albumin (f-BSA)] in the presence of moisture and physiological temperature. Coincorporation of the formaldehyde-interacting amino acid, histidine, strongly inhibits the FMAP. The purpose of this study was to test whether previous solid-state data are applicable toward the stabilization of formalinized antigens encapsulated in poly(lactide-co-glycolide) (PLGA)-based microspheres. Formaldehyde-treated bovine serum albumin (f-BSA) and BSA were selected as a model formalinized protein antigen and a nonformalinized control, respectively. As in the solid state, we found that the FMAP was dominant in the aggregation of f-BSA encapsulated in PLGA 50/50 microspheres, whereas the aggregation mechanism of encapsulated BSA was mostly converted from thiol-disulfide interchange to an acid-catalyzed noncovalent pathway. The lack of noncovalent aggregation in encapsulated f-BSA could be explained by its higher thermodynamic stability after formalinization, which inhibits protein unfolding. Targeting the FMAP, coencapsulation of histidine and trehalose successfully inhibited the aggregation of f-BSA in microspheres. By combining the use of an optimized oil-in-oil (o/o) encapsulation method, coencapsulation of histidine and trehalose, and use of low-acid-content poly(D,L-lactide) (PLA) and poly(ethylene glycol) (PEG) blends, a 2-month continuous release of f-BSA was achieved with the absence of aggregation.
authors.affiliation: Department of Pharmaceutical Sciences, The University of Michigan, Ann Arbor, Michigan 48109-1065, USA.
authors.forename: W
authors.initials: W
authors.lastname: Jiang
authors.forename: S P
authors.initials: SP
authors.lastname: Schwendeman
mesh_terms: D000818:Animals; D000951:Antigens, Neoplasm / Q000737:chemistry* / Q000648:ultrastructure; D002417:Cattle; D003692:Delayed-Action Preparations / Q000737:chemistry; D004337:Drug Carriers / Q000737:chemistry; D004355:Drug Stability; D004655:Emulsions; D005079:Excipients / Q000737:chemistry; D007700:Kinetics; D008855:Microscopy, Electron, Scanning; D008863:Microspheres; D011091:Polyesters / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D011098:Polyglactin 910 / Q000737:chemistry*; D011489:Protein Denaturation; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000276:immunology / Q000648:ultrastructure; D014612:Vaccines
chemical_list: D000951:Antigens, Neoplasm; D003692:Delayed-Action Preparations; D004337:Drug Carriers; D004655:Emulsions; D005079:Excipients; D011091:Polyesters; D014612:Vaccines; C094440:formalinized extracellular antigen; D012710:Serum Albumin, Bovine; D011098:Polyglactin 910; D011092:Polyethylene Glycols; C033616:poly(lactide)