Microencapsulated engineered Lactococcus lactis cells for heterologous protein delivery: preparation and in vitro analysis.
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.
Lawuyi
Bisi
B
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.
Chen
Hongmei
H
Afkhami
Fatemeh
F
Kulamarva
Arun
A
Prakash
Satya
S
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
D000464:Alginates; D006603:Hexuronic Acids; D020723:Glucuronic Acid; D008836:Micrococcal Nuclease