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Dr Peter Nilsson
Marcus Gry (PhD student), Ulrika Igel (PhD student), Maja Neiman (PhD student), Rebecca Rimini
(researcher), Jochen Schwenk (researcher), Ronald Sjöberg (research engineer), Mårten Sundberg (research engineer)
To analyse the specificity and quality of all purified HPA antibodies, develop and utilize 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.
Three complementary microarray formats for large-scale analysis of serum samples are under development and being utilized. 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 enable serum from very large patient cohorts to be analyzed simultaneously. In addition is also a bead-based suspension array format being utilized, with capacity for multiplexing in two dimensions, samples and antibodies.
Comparative expression profiling of microarray derived mRNA profiles and protein expression profiles from the HPR cell lines are being performed, with the aim to obtain a global comparison of the transcriptome and proteome.
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