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Alhydrogel® adjuvant, ultrasonic dispersion and protein binding: A TEM and analytical study

Lookup NU author(s): Professor Robin Harris, Dr Andrei Soliakov, Professor Rick Lewis, Professor Jeremy Lakey

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Abstract

Aluminium-based vaccine adjuvants have been in use since the 1920s. Aluminium hydroxide (alum) that is the chemical basis of Alhydrogel, a widely used adjuvant, is a colloid that binds proteins to the particular surface for efficient presentation to the immune system during the vaccination process. Using conventional TEM and cryo-TEM we have shown that Alhydrogel can be finely dispersed by ultrasonication of the aqueous suspension. Clusters of ultrasonicated aluminium hydroxide micro-fibre crystals have been produced (similar to 10-100 nm), that are significantly smaller than those present the untreated Alhydrogel (similar to 2-12 mu m). However, even prolonged ultrasonication did not produce a homogenous suspension of single aluminium hydroxide micro-fibres. The TEM images of unstained and negatively stained air-dried Alhydrogel are very similar to those obtained by cryo-electron microscopy. Visualization of protein on the surface of the finely dispersed Alhydrogel by TEM is facilitated by prior ultrasonication. Several examples are given, including some of medical relevance, using proteins of widely ranging molecular mass and oligomerization state. Even with the smaller mass proteins, their presence on the Alhydrogel surface can be readily defined by TEM. It has been found that low quantities of protein tend to cross-link and aggregate the small Alhydogel clusters, in a more pronounced manner than high protein concentrations. This indicates that complete saturation of the available Alhydrogel surface with protein may be achieved, with minimal cross-linkage, and future exploitation of this treatment of Alhydrogel is likely to be of immediate value for more efficient vaccine production. (C) 2011 Elsevier Ltd. All rights reserved.


Publication metadata

Author(s): Harris JR, Soliakov A, Lewis RJ, Depoix F, Watkinson A, Lakey JH

Publication type: Article

Publication status: Published

Journal: Micron

Year: 2012

Volume: 43

Issue: 2-3

Pages: 192-200

Print publication date: 01/02/2012

Online publication date: 21/07/2011

Acceptance date: 11/07/2011

ISSN (print): 0968-4328

ISSN (electronic): 1878-4291

Publisher: Elsevier Ltd.

URL: http://dx.doi.org/10.1016/j.micron.2011.07.012

DOI: 10.1016/j.micron.2011.07.012


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