Solubility and binding properties of PEGylated lysozyme derivatives with increasing molecular weight on hydrophobic-interaction chromatographic resins

Müller, Egbert; Josić, Djuro; Schröder, Tim; Moosmann, Anna

doi:10.1016/j.chroma.2010.05.016

Cited by 31 publications

(29 citation statements)

References 36 publications

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“…In addition, a small difference in the linker structure can result in different presentations of the metallacarborane moiety on the protein surface, thus affecting the affinity of the conjugate to the HIC column. A similar phenomenon for lysozyme–polyethylene glycol (PEG) conjugates was observed 31. In the cited case, small changes in isoelectric points, surface charge distributions, or relative hydrophobicities, in addition to different locations of PEGylation on the protein surface, were proposed as possible reasons for the observable differences in chromatographic mobility.…”

Section: Resultssupporting

confidence: 57%

Synthesis of Lysozyme–Metallacarborane Conjugates and the Effect of Boron Cluster Modification on Protein Structure and Function

et al. 2015

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Two complementary methods, "in solution" and "in solid state", for the synthesis of lysozyme modified with metallacarborane (cobalt bis(dicarbollide), Co(C2 B9 H11 )2 (2-) ) were developed. As metallacarborane donors, oxonium adducts of cobalt bis(dicarbollide) and 1,4-dioxane or tetrahydropyran were used. The physicochemical and biochemical properties of the obtained lysozyme-metallacarborane conjugates were studied for changes in secondary and tertiary structure, aggregation behavior, and biological activity. Only minor changes in primary, secondary, and tertiary protein structure were observed, caused by the single substitution of metallacarborane on lysozyme. However, the modification produced significant changes in lysozyme enzymatic activity and a tendency toward time- and temperature-dependent aggregation.

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Section: Resultssupporting

confidence: 57%

Synthesis of Lysozyme–Metallacarborane Conjugates and the Effect of Boron Cluster Modification on Protein Structure and Function

et al. 2015

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“…This has been achieved by use of buffers in which PEGylated proteins can be fully dissolved. For instance, good PEGamer resolution was achieved when PEG-lysozyme conjugates were separated in sodium chloride, by a still unknown supporting mechanism, compared with the separation achieved in ammonium sulfate, in which lower solubility of the same proteins was observed [21]. Because the changes in the hydrophobic properties of PEGylated proteins are difficult to predict, and chain lengths also affect column-binding capacity, and because of the apparent increase in molecular weight conferred by solubilized conjugates, separation conditions for this technique must be determined empirically.…”

Section: Separation and Purification Of Pegylated Conjugatesmentioning

confidence: 95%

Advances and trends in the design, analysis, and characterization of polymer–protein conjugates for “PEGylaided” bioprocesses

2012

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In addition to their use as therapeutics and because of their enhanced properties, PEGylated proteins have potential application in fields such as bioprocessing. However, the use of PEGylated conjugates to improve the performance of bioprocess has not been widely explored. This limited additional industrial use of PEG-protein conjugates can be attributed to the fact that PEGylation reactions, separation of the products, and final characterization of the structure and activity of the resulting species are not trivial tasks. The development of bioprocessing operations based on PEGylated proteins relies heavily in the use of analytical tools that must sometimes be adapted from the strategies used in pharmaceutical conjugate development. For instance, to evaluate conjugate performance in bioprocessing operations, both chromatographic and non-chromatographic steps must be used to separate and quantify the resulting reaction species. Characterization of the conjugates by mass spectrometry, circular dichroism, and specific activity assays, among other adapted techniques, is then required to evaluate the feasibility of using the conjugates in any operation. Correct selection of the technical and analytical methods in each of the steps from design of the PEGylation reaction to its final engineering application will ensure success in implementing a "PEGylaided" process. In this context, the objective of this review is to describe technological and analytical trends in developing successful applications of PEGylated conjugates in bioprocesses and to describe potential fields in which these proteins can be exploited.

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“…In order to achieve a welldefined product, further separation, mostly by use of different chromatographic methods, is necessary. MALDI-TOF and MALDI-TOF/TOF have proved to be reliable methods for the thorough analysis of product size and composition [128].…”

Section: Clotting Factor IXmentioning

confidence: 99%

Therapeutic plasma proteins – application of proteomics in process optimization, validation, and analysis of the final product

et al. 2011

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An overview is given on the application of proteomic technology in the monitoring of different steps during the production of therapeutic proteins from human plasma. Recent advances in this technology enable the use of proteomics as an advantageous tool for the validation of already existing processes, the development and fine tuning of new production steps, the characterization and quality control of final products, the detection of both harmful impurities and modifications of the therapeutic protein and the auditing of batch-to-batch variations. Further, use of proteomics for preclinical testing of new products, which can be either recombinant or plasma-derived, is also discussed.

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Solubility and binding properties of PEGylated lysozyme derivatives with increasing molecular weight on hydrophobic-interaction chromatographic resins

Cited by 31 publications

References 36 publications

Synthesis of Lysozyme–Metallacarborane Conjugates and the Effect of Boron Cluster Modification on Protein Structure and Function

Synthesis of Lysozyme–Metallacarborane Conjugates and the Effect of Boron Cluster Modification on Protein Structure and Function

Advances and trends in the design, analysis, and characterization of polymer–protein conjugates for “PEGylaided” bioprocesses

Therapeutic plasma proteins – application of proteomics in process optimization, validation, and analysis of the final product

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