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A new size and shape controlling method for producing calcium alginate beads with immobilized proteins
Department of Biotechnology, Grad. School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, 565-0871, Osaka, Japan;
Department of Biotechnology, Grad. School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, 565-0871, Osaka, Japan;
Department of Applied Physics, Grad. School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, 565-0871, Osaka, Japan
Department of Applied Physics, Grad. School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, 565-0871, Osaka, Japan
Department of Applied Physics, Grad. School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, 565-0871, Osaka, Japan
Department of Applied Physics, Grad. School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, 565-0871, Osaka, Japan
- 1 Department of Biotechnology, Grad. School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, 565-0871, Osaka, Japan;
- 2 Department of Biotechnology, Grad. School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, 565-0871, Osaka, Japan;
- 3 Department of Applied Physics, Grad. School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, 565-0871, Osaka, Japan
- 4 Department of Applied Physics, Grad. School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, 565-0871, Osaka, Japan
- 5 Department of Applied Physics, Grad. School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, 565-0871, Osaka, Japan
- 6 Department of Applied Physics, Grad. School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, 565-0871, Osaka, Japan
Journal of Biomedical Science and Engineering·Volume 02 (2009)·Pages 287–293·Published 17 September 2009·DOI10.4236/jbise.2009.25043
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Abstract
A method for producing size- and shape-con-trolled calcium alginate beads with immobilized proteins was developed. Unlike previous cal-cium alginate bead production methods, pro-tein-immobilized alginate beads with uniform shape and sizes less then 20 micrometers in diameter could successfully be produced by using sonic vibration. BSA and FITC-conjugated anti-BSA antibodies were used to confirm pro-tein immobilization in the alginate beads. Pro-tein diffusion from the beads could be reduced to less than 10% by cross-linking the proteins to the alginate with 1-ethyl-3-(3-dimethylamino-propyl)carbodiimide (EDC) and N-hydroxysul-fosuccinimide (NHSS). The calcium alginate beads could also be arranged freely on a slide glass by using a femtosecond laser.
KeywordsCalcium Alginate BeadsSize Controllable Production MethodProtein Immobilized BeadsFemtosecond LaserLaser Manipulation
- Mizukami, A., Nagamori, E., Takakura, Y., Matsunaga, S., Kaneko, Y., Kajiyama, S., Harashima, S., Kobayashi, A., and Fukui, K., (2003) Transformation of yeast using calcium algi-nate microbeads with surface-immobilized chromosomal DNA, Biotechniq., 35, 734–736, 738–740.
- Liu, H., Kawabe, A., Matsunaga, S., Murakawa, T., Mizukami, A., Yanagisawa, M., Nagamori, E., Harashima, S., Kobayashi, A., and Fukui, K., (2004) Obtaining transgenic plants using the calcium alginate beads method, J. Plant Res., 117, 95–99.
- Sone, T., Nagamori, E., Ikeuchi, T., Mizukami, A., Takakura, Y., Kajiyama, S., Fukusaki, E., Harashima, S., Kobayashi, A., and Fukui K., (2002) A novel gene delivery system in plants with calcium alginate micro-beads, J. Biosci. Bioeng., 94, 87–91.
- Higashi, T., Nagamori, E., Sone, T., Matsunaga, S. and Fukui, K. (2004) A novel transfection method for mammalian cells using calcium alginate microbeads, J. Biosci. Bioeng., 97, 191–195.
- Gray, C. J. and Dowsett, J., (1988) Retention of insulin in alginate gel beads, Biotech. Bioeng., 31, 607–612.
- Ko, C., Dixit, V., Shaw, W., and Gitnick, G., (1995) In vitro slow release profile of endothelial cell growth factor immobi-lized within calcium alginate microbeads, Artif. Cells Blood Substit Immobil. Biotechnol., 23, 143– 151.
- Smidsr?d, O. and Skj?k-Braek, G., (1990) Alginate as immobi-lization matrix for cells, Trends Biotechnol., 8, 71–78.
- Puolakkainen, P. A., Ranchalis, J. E., Gombotz, W. R., Hoff-man, A. S., Mumper, R. J., and Twardzik, D. R., (1994) Novel delivery system for inducing quiescence in intestinal stem cells in rats by transforming growth factor beta 1, Gastroenterology, 107, 1319–1326.
- Singh, O. N. and Burgess, J., (1989) Characterization of albu-min-alginic acid complex coacervation, J. Pharm. Pharmacol., 41, 670–673.
- Robitaille, R., Pariseau, J. F., Leblond, F. A., Lamoureux, M., Lepage, Y., and Hallé, J. P., (1999) Studies on small (<350 microm) alginate-poly-L-lysine microcapsules. III. Biocom-patibility of smaller versus standard microcapsules, J. Biomed. Mater. Res., 44, 116–120.
- Poncelet, D. and Neufeld R. J., (1989) Shear breakage of nylon membrane microcapsules in a turbine reactor, Biotechnol. Bioeng., 5, 95–103.
- Sakai, S., Ono, T., Ijima, H., and Kawakami, K., (2000) Syn-thesis and transport characterization of alginate/ aminopro-pyl-silicate/alginate microcapsule: application to bioartificial pancreas, Biomater., 22, 2827–2834.