Protection of the enzyme l -asparaginase during lyophilisation—a molecular modelling approach to predict required level of lyoprotectant

Protection of the enzyme l -asparaginase during lyophilisation—a molecular modelling approach... Many novel therapeutic agents are proteins and peptides which need stabilisation due to their inherent instability in aqueous solution. Freeze-drying is an established method for protein stabilisation, although the use of additives is often necessary in order to preserve protein structure and activity during lyophilisation itself. The molecular interactions between protein and protective additive are as yet unclear. In this study, we examined the use of a range of saccharide additives to stabilise the model multi-subunit enzyme l -asparaginase during lyophilisation, assessed post-drying enzyme activity and quaternary structure, and related the extrapolated levels of additive necessary to provide full stabilisation to the theoretical levels predicted from an existing hypothesis using molecular modelling. It was found that each of the saccharides tested here displayed similar levels of protection towards l -asparaginase under the conditions used. Amounts of additive required to give full stabilisation to the enzyme were extrapolated from the activity data and were found to be in good agreement with theoretical amounts calculated from molecular modelling studies. Our data suggest that the existing hypothesis may be relevant to the prediction of optimum levels of lyoprotectant for the freeze-drying of proteins. However, further studies would be necessary in order to obtain a full picture of protein-additive interactions at the molecular level. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png International Journal of Pharmaceutics Elsevier

Protection of the enzyme l -asparaginase during lyophilisation—a molecular modelling approach to predict required level of lyoprotectant

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Publisher
Elsevier
Copyright
Copyright © 1999 Elsevier Science B.V.
ISSN
0378-5173
D.O.I.
10.1016/S0378-5173(99)00163-5
Publisher site
See Article on Publisher Site

Abstract

Many novel therapeutic agents are proteins and peptides which need stabilisation due to their inherent instability in aqueous solution. Freeze-drying is an established method for protein stabilisation, although the use of additives is often necessary in order to preserve protein structure and activity during lyophilisation itself. The molecular interactions between protein and protective additive are as yet unclear. In this study, we examined the use of a range of saccharide additives to stabilise the model multi-subunit enzyme l -asparaginase during lyophilisation, assessed post-drying enzyme activity and quaternary structure, and related the extrapolated levels of additive necessary to provide full stabilisation to the theoretical levels predicted from an existing hypothesis using molecular modelling. It was found that each of the saccharides tested here displayed similar levels of protection towards l -asparaginase under the conditions used. Amounts of additive required to give full stabilisation to the enzyme were extrapolated from the activity data and were found to be in good agreement with theoretical amounts calculated from molecular modelling studies. Our data suggest that the existing hypothesis may be relevant to the prediction of optimum levels of lyoprotectant for the freeze-drying of proteins. However, further studies would be necessary in order to obtain a full picture of protein-additive interactions at the molecular level.

Journal

International Journal of PharmaceuticsElsevier

Published: Oct 5, 1999

References

  • Factors affecting short-term and long-term stabilities of proteins
    Arakawa, T.; Prestrelski, S.J.; Kenney, W.C; Carpenter, J.F
  • Rational design of stable lyophilized protein formulations: some practical advice
    Carpenter, J.F.; Pikal, M.J.; Chang, B.S.; Randolph, T.W.
  • The adverse effect of glycation of human serum albumin on its preservative activity in the freeze-drying and accelerated degradation of alkaline phosphatase
    Ford, A.W.; Allahiary, Z.
  • Stabilizing effect of amphiphilic excipients on the freeze-thawing and freeze-drying of lactate dehydrogenase
    Izutsu, K.; Yoshioka, S.; Terao, T.

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