title: Novel, high throughput method to study in vitro protein release from polymer nanospheres.
abstract: Controlled delivery of therapeutic protein drugs using biodegradable polymer carriers is a desired characteristic that enables effective, application-specific therapy and treatment. Previous studies have focused on protein delivery from polymers using conventional "one-sample-at-a-time" techniques, which are time-consuming and costly. In addition, many therapeutic proteins are in limited supply and are expensive, so it is desirable to reduce sample size for design and development of delivery devices. We have developed a rapid, high throughput technique based on a highly sensitive fluorescence-based assay to detect and quantify protein released from polyanhydrides while utilizing relatively small amounts of protein (approximately 40 microg). These studies focused on the release of a model protein, Texas Red conjugated bovine serum albumin, from polyanhydride copolymers based on sebacic acid (SA) and 1,6-bis(p-carboxyphenoxy)hexane (CPH). The protein release profiles were assessed simultaneously to investigate the effect of polymer device geometry (nanospheres vs films), polymer chemistry, and pH of the release medium. The results indicated that the nanosphere geometry, SA-rich chemistries, and neutral pH release medium led to a more rapid release of the protein compared to the film geometry, CPH-rich chemistries, and acidic pH release medium, respectively. This high throughput fluorescence-based method can be readily extended to study release kinetics for other proteins and polymer systems.
authors.affiliation: Department of Chemical and Biological Engineering, Iowa State University, Ames, Iowa 50011, USA.
authors.forename: L K
authors.initials: LK
authors.lastname: Petersen
authors.forename: C K
authors.initials: CK
authors.lastname: Sackett
authors.forename: B
authors.initials: B
authors.lastname: Narasimhan
mesh_terms: D000818:Animals; D001798:Blood Proteins / Q000737:chemistry; D002417:Cattle; D020650:Combinatorial Chemistry Techniques / Q000379:methods*; D004337:Drug Carriers / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D054118:Nanospheres / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry*; D012710:Serum Albumin, Bovine; D013499:Surface Properties; D014966:Xanthenes / Q000737:chemistry
chemical_list: D001798:Blood Proteins; D004337:Drug Carriers; D005456:Fluorescent Dyes; D011108:Polymers; D011506:Proteins; D014966:Xanthenes; D012710:Serum Albumin, Bovine; C034657:Texas red