Modification of surfaces and proteins by attaching poly(ethylene glycol) (PEG) is an important technique for controlling the properties of these materials. Our goal is to examine the possible combination of these effects by linking proteins to surfaces via a PEG spacer or tether. In the present work we have coupled alkaline phosphatase (as a model protein) to porous glass by means of PEG spacers, and we have compared the activity and operational sta bility of the PEG-bound enzyme to free enzyme and to enzyme immobilized by a conventional, short urea linkage. Significantly, the PEG-bound enzyme differs little in its catalytic properties from free enzyme, indicating that the bound enzyme extends into solution and is in essence free of the surface.
Karr, L.J., S.G. Shafer, J.M. Harris, J.M. Van Alstine and R.S. Snyder.J. Chrom., 354:269-282 (1986).
3.
Karr, L.J., J.M. Van Alstine, R.S. Snyder, S.G. Shafer and J.M. Harris.J. Chrom., 442:219-227 (1988).
4.
Yoshinaga, K. , S.G. Shafer and J.M. Harris.J. Bioact. Compatible Polym., 2:49-56 (1987).
5.
Harris, J.M., K. Yoshinaga, M.S. Paley and M.R. Herati. "New Activated PEG Derivatives for Affinity Partitioning ," Advances in Separations Using Aqueous Phase Systems in Cell Biology and Biotechnology. D. Fisher and I. A. Sutherland, eds. London:Plenum (1988) in press.
6.
Harris, J.M. and K. Yoshinaga.J Bioact. Compatible Polym. , 4:17-24 (1989).
7.
(a) MacEwen, E.G., R. Rosenthal, R. Matus, A.T. Viau and A. Abuchowski, Cancer, 59:2011-2015 (1987);
8.
(b) A.T. Viau, A. Abuchowski, S. Greenspan, F.F. Davis, J. Free Radicals Biol. Med., 2:283-288 (1986).
9.
Takahashi, K. , A. Ajima, T. Yoshimoto, M. Okada, A. Matsushima, Y. Tamaura and Y. Inada.J. Org. Chem. , 50:3414 (1985).
Graham, N.B. in Hydrogels in Medicine and Pharmacy, Vol. II. N. A. Peppas, ed. Boca Raton, Florida:CRC Press, Chap. 4 (1987).
14.
Merrill, E.W., E.W. Salzman, K.A. Dennison, S.-W. Tay and R.W. Pekala in Progress in Artifical Organs-1985. Y. Nose, C. Kjellstrand, and P. Ivanovich, eds. ISAO Press: Cleveland, pp. 909-912 (1986).
15.
Gombotz, W.R. , W. Guanghui and A.S. Hoffman.J. Appl. Polym. Sci., in press.
16.
Bamberger, S.B., J.M. Van Alstine, J.M. Baird, J.M. Harris and D.E. Brooks. "Demixing of Aqueous Polymer Two-Phase Systems in the Absence of Gravity," Sep. Sci. Tech., 23:17-34 (1987).
17.
Harris, J.M. , D.E. Brooks, J.F. Boyce, R.S. Snyder and J.M. Van Alstine in "Dynamic Aspects of Polymer Surfaces ." J. D. Andrade, ed. Plenum, pp. 111-119 (1988).
18.
Bayer, E., M. Dengler and B. Hemmasi.Int. J Peptide Res., 25:178 (1985).
19.
Becker, H., H. Lucas, J. Maul, R. Pillai, H. Anzinger and M. Mutter.Makromol. Chem. Rapid Comm., 3:217 (1982).
20.
Zalipsky, S. , F. Albericio and G. Barany in Peptides: Structure and Function. C. M. Deber, V. J. Hruby and K. D. Kopple, eds. Pierce Chemical Co. , p. 257 ( 1985).
21.
Grainger, D.W. , S.W. Kim and J. Feijen.J. Biomed. Mat. Res., 22:231-249 (1988).
22.
S. Winters, Doctoral Thesis, University of Utah, 1987.
23.
Tsuchida, E. , H. Nishide, N. Shimidzu, A. Yamada, M. Kaneko and Y. Kurimura.Makromol. Chem. Rapid Commun., 2:621 ( 1981).
24.
Torres, J.L. , R. Guzman, R.G. Carbonell and P.K. Kilpatrick.Anal. Biochem. , 171:411-418 (1988).
25.
Drevin, I. and B. Johansson.J Chrom., 295:210 (1984).
26.
Carr, P.W.Immobilized Enzymes in Analytical and Clinical Chemistry. New York:Wiley-Interscience (1980 ).
27.
Leyden, D. E., ed. Silanes, Surfaces and Interfaces. New York: Gordon and Breach (1986).
28.
Sedaghat-Herati, R. unpublished results.
29.
Gölander, C.G. and E. Kiss.J Coll. Int. Sci., 121:240 (1988).
30.
Coleman, J.E. and P. Gettins.Advan. Enzymol., 55:351 (1983).
