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Dr Peter Nilsson
Paula Borg (technician), Marcus Gry (PhD student), Rebecca Rimini
(researcher), Jochen Schwenk (post doc), Ronald Sjöberg (research engineer), Mårten Sundberg (research engineer), Annelie Walden (research engineer)
To analyse the specificity and quality of all purified HPA antibodies, develop microarray-based methodologies for large scale analysis of serum samples with HPA antibodies and to analyse the transcriptome of the HPR cell lines.
Methodology for microarray based analysis of antibody specificity has been developed, where all purified antibodies are analyzed on protein arrays with immobilized PrESTs. Each microarray is divided into 14 distinct areas (each containing 384 PrESTs) with a multi well device, enabling the analysis of 14 antibodies simultaneously. The antibodies are detected through a fluorescently labeled secondary antibody. A specificity plot is generated for each antibody, where the signal from the binding to its antigen is compared to the unspecific binding to all the other PrESTs. A dual color system is used in order to verify the presence of the spotted PrESTs.
Two complementary microarray formats for large-scale analysis of serum samples are under development. These are antibody microarrays with the possibility for simultaneous analysis of large amounts of analytes with high sensitivity and the reverse phase serum microarrays which enables serum from very large patient cohorts to be analyzed simultaneously. In addition is also a bead-based suspension array format being evaluated.
Comparative expression profiling of microarray derived mRNA profiles and protein expression profiles from the HPA cell lines are being performed, with the aim to obtain a global comparison of the transcriptome and proteome.
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