Evaluation of temporal aggregation processes using spatial intensity distribution analysis

Zahra Rattray, Egor Zindy, Karen M. Buzza, Alain Pluen

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Small proteinaceous oligomeric species contribute to the formation of larger aggregates, a phenomenon that is of direct relevance to the characterization of protein aggregation in biopharmaceuticals and understanding the underlying processes contributing to neurodegenerative diseases. The ability to monitor in situ oligomerization and aggregation processes renders imaging and image analysis an attractive approach for gaining a mechanistic insight into early processes contributing to the formation of larger aggregates in disease models and biologics. The combination of image analysis tools enables the detection of both oligomeric and larger aggregate subtype in contrast to conventional kinetic-based approaches that lack the ability to resolve dimers from monomeric moieties in samples containing mixed populations. In this chapter, we describe the process for confocal time series image acquisition for monitoring the in situ loss of monomers, and the subsequent analysis pipeline using spatial intensity distribution analysis (SpIDA) to evaluate oligomer content.
Original languageEnglish
Title of host publicationProtein Self-Assembly
Subtitle of host publicationMethods and Protocols
EditorsJennifer J. McManus
Place of PublicationNew York, NY
PublisherSpringer
Pages141-155
Number of pages15
ISBN (Print)9781493996773, 9781493996780
DOIs
Publication statusPublished - 23 Sep 2019

Publication series

NameMethods in Molecular Biology
Volume2039

Keywords

  • protein aggregation
  • image analysis
  • microscopy

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