title: Microencapsulated engineered Lactococcus lactis cells for heterologous protein delivery: preparation and in vitro analysis.
abstract: This article demonstrates the potential of encapsulated, engineered Lactococcus lactis as a vehicle for the oral delivery of therapeutic proteins. Using alginate-poly-l-lysine-alginate membrane-encapsulated L. lactis engineered to secrete the reporter protein Staphylococcal aureus nuclease, we show comparable viability and protein secretion between free and immobilized cells. After 12 h, microcapsules with a cell density of 4.8 x 10(5) colony forming unit (CFU) ml(-1) grew to 2.2 x 10(8) CFU ml(-1) and released 0.24 arbitrary unit (AU) ml(-1) of nuclease, producing similar results as free cells, which grew from 3.4 x 10(5) to 1.9 x 10(8) CFU ml(-1) and secreted 0.21 AU ml(-1) of nuclease. Moreover, encapsulated cells at a density of 4.4 x 10(7) CFU ml(-1) grew to 2.2 x 10(10) CFU ml(-1) in 12 h and secreted 15.3 AU ml(-1) of nuclease although 3.1 x 10(7) CFU ml(-1)of free cells reached only 2.3 x 10(9) CFU ml(-1) and released 5.6 AU ml(-1) of nuclease. We also show the sustained stability of the microcapsules during storage at 4 degrees C over 8 weeks.
authors.affiliation: Biomedical Technology and Cell Therapy Research Laboratory, Department of Biomedical Engineering and Artificial Cells and Organs Research Center, Faculty of Medicine, McGill University, 3775 University Street, Montreal, Quebec, Canada.
authors.forename: Bisi
authors.initials: B
authors.lastname: Lawuyi
authors.forename: Hongmei
authors.initials: H
authors.lastname: Chen
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Afkhami
authors.forename: Arun
authors.initials: A
authors.lastname: Kulamarva
authors.forename: Satya
authors.initials: S
authors.lastname: Prakash
mesh_terms: D000464:Alginates / Q000737:chemistry; D001709:Biotechnology / Q000379:methods*; D002478:Cells, Cultured; D004339:Drug Compounding; D004591:Electrophoresis, Polyacrylamide Gel; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D013294:Lactococcus lactis / Q000378:metabolism*; D008836:Micrococcal Nuclease / Q000378:metabolism; D008863:Microspheres; D015202:Protein Engineering / Q000379:methods*; D013234:Stem Cells; D013696:Temperature; D013997:Time Factors
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D020723:Glucuronic Acid; D008836:Micrococcal Nuclease