Validating sec-ls aggregation methods -

In this commentary, we, as representatives of EIP-PCS, review the current state of methods for analysis of protein aggregates. Moreover, we elaborate on why these...

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The EIP represents companies, institutes and professionals involved in research, development, testing, validation, application, production or marketing of immunogenicity assessment tools, as well as those servicing the biotechnology community.

Recently, Philo reviewed AUC as a method for characterizing non-particulate, soluble protein aggregates, including its strengths and weaknesses Structural properties of monoclonal antibody aggregates induced by freeze-thawing and thermal stress. Micelle-associated protein in epoetin formulations: Extended characterization data, if available, may then help to establish a relationship between the presence and nature of aggregates in a protein pharmaceutical on the one hand and immunogenicity data emerging from use of the product in humans on the other.

Broad size range; no stationary phase; quantification; can be combined with serial online detectors.

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Since small proteins have a relatively large diffusion coefficient, they will return more rapidly to the fast streamlines of the laminar channel flow than large proteins. The use of asymmetrical flow field-flow fractionation in pharmaceutics and biopharmaceutics. In order to minimize particle problems as much as possible, it is recommended to use harmonized inspection procedures and acceptance criteria throughout development especially for release testing and to evaluate the performance of the visual inspection method before manufacture of GMP batches.

Open Access This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author s and source are credited. In these cases, sterile filtration is often not effective because insoluble protein particles may reform over time.

As each method covers different aggregate characteristics, the results obtained with a particular method are strictly linked to that method. The addition to the eluent of fluorescent dyes that bind to denatured protein and so inducing an enhancement of the fluorescence allows detection of very low amount of denatured protein

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  • The combination of SEC with light-scattering (SEC/LS) detection was The Arrhenius approach over-predicted the...
  • Method validation investigated parameters such as the range, The results demonstrate the validity...
  • Strategies for the Assessment of Protein Aggregates in Pharmaceutical Biotech Product...
  • PDF | Although size exclusion chromatography (SEC) has been, and...
  • How can we validate AUC methods for non-covalent aggregates given...

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Validating sec-ls aggregation methods

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Focus on aggregation: types, causes, characterization, and impact John Philo V.P. . nearly always ask for cross-validation of SEC methods by orthogonal methods; Methods typically used to cross-check SEC analytical 0 5 10 15 20...