|
|
 |
 |
Dr Peter Nilsson
Burcu Ayoglu (PhD-student), Kimi Drobin (research engineer), Anna Gundberg (technician), Anna Häggmark (PhD-student), Ulrika Igel (PhD-student), Maja Neiman (PhD-student),
Jochen M. Schwenk (researcher), Ronald Sjöberg (research engineer) and 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 plasma/serum samples with HPA antibodies.
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.
Several complementary microarray formats for systematic analysis of plasma/serum samples are being utilized and under constant development. The PrEST-arrays have been implemented for systematic antigen-based plasma profiling for the screening of new autoimmunity components. The 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 are both utilizing in-house produced planar microarrays. The main platform for systematic antibody-based plasma profiling is the suspension bead array format, with capacity for multiplexing in two dimensions, enabling the simultaneous profiling of 384 antibodies on 384 samples, see also Module 12: Plasma profiling.
home | about hpr - organization | module 5
|
|
|
|
|
|
