Summary
The production of pharmaceutical grade proteins using recombinant DNA technology necessitates a commitment on the part of the manufacturer to analyze critically these biomolecules with respect to both identity and purity. Routine analysis of recombinant proteins currently involves techniques relating to the evaluation of specific activity, homogeneity, amino acid composition, peptide maps, and N-terminal and C-terminal analyses. Confirmation of the fidelity of a recombinant protein’s primary structure is the result of sequence analysis at both the protein and the DNA levels, and implies a shared responsibility between the protein chemist and molecular biologist. Once generated, the information serves as a reference standard for progressive lots of production material. The ability to perform certain analyses is in part a reflection of the limitations of current technology. As recombinant proteins become more complex, e.g., hetero-oligomers, post-translationally modified, larger mass, constraints as to the type and degree of analysis are encountered. This paper describes areas of routine investigation for recombinant proteins and highlights some practical limitations of their analysis.
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References
Commercial Biotechnology: An International Analysis (Washington, D.C.: U.S. Congress Office of Technology Assessment, OTA-BA-218, January, 1984 ).
Revel, M. (1984) in Antiviral Drugs and Interferon: The Molecular Basis of Their Activity. (Y. Becker, ed.) Martinus Nishoff Publishing: Boston, pp. 357–433.
Pennica, D. et al. (1983) Nature 301, 214–221.
Cate, R.L. et al. (1986) Cell 45, 685–698.
Federal Register (1986) 51, 23302.
The United States Pharmacopeia/The National Formulary. (1985) United States Pharmacopeia Convention, Inc.: Rockville, MD.
Taniguchi, T. et al. (1983) Nature 302, 305–310.
Maniatis T. Fritsch, E.F., and Sambrook, J. (1982) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Maxam, A.M. and Gilbert, W. (1980) Methods in Enzymol. 65, 499–560.
Winkler, M.E. et al. (1985) Biotechnology 3, 990–998.
Wood, W.I. et al. (1984) Nature 312, 330–337.
Kaufman, R.J. and Sharp, P.A. (1982) J. Mol. Biol. 159, 601–621.
Gitschier, J. et al. (1984) Nature 312, 326–330.
Hunkapillar, M.W. et al. (1984) Science 226, 304–311.
Lahm, H.-W., and Stein, S. (1985) J. Chromatogr. 326, 357–361.
Liang, S.-M. et al. (1985) Biochem. J. 229, 429–439.
Rose, K. et al. (1986) submitted.
Ambler, R.P. (1972) Methods Enzymol. 25B, 143–154.
Rose, K. et al. (1983) Biochem. J. 215, 273–277.
Stark, G.R. (1972) Methods Enzymol. 25B, 369–384
Browning, J.L. et al. (1986) Anal. Biochem. 155, 123128.
Biemann, K.; Carr, S.A., and Roberts, G.D.; this volume.
Carr, S.A. and Biemann, K. (1984) Methods in Enzymol. 106, 29–58.
Hunt, D.F. et al. (1986) Proc. Natl. Acad. Sci. 83, 6233–6237.
Wang, A. et al. (1984) Science 24, 1431–1433.
Liang, S.-7-M:M. et al. (1986) J. Biol. Chem. 261, 334–337.
Bewley, T.A. (1985) in Therapeutic Agents Produced by Genetic Engineering: Quo Vadis? (A. Joyeaux, G. Leygue, M. Morre, R. Roncucci, P.H. Schmelck, eds.) “Quo Vadis?” Symposium, Sanofi Group, May 29–30, 1985. Toulouse-Laberge, France.
Liang, C.-M. et al. (1985) Biochem. Biophys. Res. Comm. 128, 171–178
Petricciani, J.C. (1979) Adv. Exp. Med. Biol. 118, 921.
Nick Translation Kit, #81605B, Bethesda Research Laboratories.
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Mattaliano, R.J., Rosa, J.J., Foeller, C., Woodard, J.P., Bertolini, M.J. (1987). Analysis of Recombinant Proteins — Current Trends and Practical Limits in Analytical Stringency. In: Walsh, K.A. (eds) Methods in Protein Sequence Analysis · 1986. Experimental Biology and Medicine, vol 14. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59259-480-1_6
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DOI: https://doi.org/10.1007/978-1-59259-480-1_6
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