Fourth International Symposium on Neuroacanthocytosis

Abstracts from the Fourth International Symposium on Neuroacanthocytosis

July 1-2, 2008
London and Oxford

Chairs: Prof. Kailash Bhatia, MD, FRCP, Institute of Neurology, University College London; Prof. Anthony P. Monaco, MD, PhD, Wellcome Trust Centre for Human Genetics, University of Oxford

Organizers: Antonio Velayos-Baeza, PhD; Susanne Schneider, MD; Glenn Irvine

5-1 Production of antibodies against human VPS13 proteins
A. Velayos Baeza, and A. P. Monaco
Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UK

The Advocacy for Neuroacanthocytosis Patients awarded us a grant in 2007 to develop new tools for the analysis of chorein and other VPS13 proteins. Two members of the VPS13 gene family have an associated disorder: Chorea-Acanthocytosis (ChAc) for VPS13A and Cohen syndrome for VPS13B. One of the main problems in the research on the molecular biology aspects of ChAc is the difficulty in obtaining good antibodies able to detect chorein, the protein encoded by VPS13A. The same applies to the rest of the VPS13 proteins (B, C and D). These antibodies are needed for the detection of the endogenous proteins not only for basic research but also for diagnostic purposes. A western-blot-based diagnostic assay is already available for ChAc, where an antibody against the N-terminal region of chorein is used. With the new antibodies we want to improve this assay and, hopefully, develop a similar one for Cohen syndrome. The approach we have followed in this project consists on over-expression of six or more different fragments for each VPS13 protein and purification of those protein fragments showing a good expression level. The purified proteins are then used as antigens to develop antibodies in different species, allowing the generation of a panel of antibodies that could be used in different combinations for a number of applications. Here we are presenting the results obtained in the cloning and over-expression of the VPS13 protein fragments as well as the initial characterisation of the first antisera obtained against six of these fragments. The next steps in the project will also be outlined.