Affinity proteomics - antibody microarray-based high throughput proteome analysis

Research projects and aims

The focus of the affinity proteomic group is to develop recombinant antibody-based microarrays for high-throughput (disease) proteomics, with a particular focus on oncoproteomics.

Technology development: We have adopted a cross-disciplinary approach to develop a state-of-the-art recombinant scFv antibody microarray technology platform by in parallell addressing all seven key array issues, 

• content (e.g. design, format, range of specificities)
• solid support (e.g. design, biocompatibility, blocking)
• array format (e.g. micro vs nano)
• array fabrication (e.g. dispensing, self-addressing)
• sample (e.g. format, labelling, aq-soluble proteins vs membrane proteins)
• analytical read-out (e.g. label-dependent vs label-free, sensitivity)
• data handling (e.g. normalization, bioinformatics)

To date, we have successfully developed the second generation of a high-performing recombinant scFv antibody microarrays for rapid, multiplexed protein expression profiling of non-fractionated, biotinylated complex proteomes, such as human sera, in a highly specific (pM to fM range), sensitive and selective manner.

Applications: We have performed differential protein expression profiling of a number of clinical samples, with a particular focus on oncoproteomics. We can target complex proteomes, such as whole serum, plasma, tissue extracts, urine, cell lysates, while consuming minute amounts of the sample (µL scale). Some of our efforts include;

• Serum protein profiling of breast cancer
• Serum protein profiling of pancreatic adenoma carcinoma
• Tissue extract profiling of gastric adenoma carcinoma
• Serum protein profiling of multiforma glioblastoma
• Serum protein profiling various inflammatory conditions (e.g. systemic lupus erythematosus, systemic sclerosis, pre-eclampsia)

Selected references

• Wingren C and Borrebaeck CAK (2007) Progress in miniaturization of protein arrays - a step closer to high-density nanoarrays. Drug Discov Today 12, 813-819.
• Borrebaeck CAK and Wingren C (2007) High-throughput proteomics using antibody microarrays: an update. Expert Rev Mol Diagn 7, 673-686.
• Ingvarsson J, Larsson A, Sjöholm AG, Truedsson L, Jansson B, Borrebaeck CAK and Wingren C (2007) Design of Recombinant Antibody Microarrays for Serum Protein Profiling: Targeting of Complement Proteins. J Proteome Res 6, 3527-3536.
• Wingren C, Ingvarsson J, Dexlin L, Szul D and Borrebaeck CAK (2007) Design of recombinant antibody microarrays for complex proteome analysis: Choice of sample labeling-tag and solid support. Proteomics 7, 3055-3065.
• Ghatnekar-Nilsson S, Dexlin L, Wingren C, Montelius L and Borrebaeck CAK (2007) Design of atto-vial based recombinant antibody arrays combined with a planar wave guide detection system. Proteomics 7, 540-547.
• Zhou Y, Xu H, Dahlin A, Vallkil J, Borrebaeck CAK, Wingren C, Liedberg B and Höök F. (2007) Quantitative interpretation of of gold nanoparticle-based bioassays designed for detection of immuno-complex formation. Biointerphases 2, 6-15.
• Wingren C and Borrebaeck CAK (2007) Antibody microarray-based proteomics. Bioforum Europe 5, 34-35.
• Ellmark P, Ingvarsson J, Carlsson A, Lundin SB, Wingren C and Borrebaeck CAK (2006) Identification of protein expression signatures associated with H. pylori infection and gastric adenocarcinoma using recombinant antibody microarrays. Mol Cell Proteomics 5, 1638-1646.
• Wingren C, Steinhauer C, Ingvarsson J, Persson E, Larsson K and Borrebaeck CAK. (2005) Microarrays based on affinity-tagged scFv antibodies: sensitive detection of analyte in complex proteomes. Proteomics 5, 1281-1291.

Contact information

Dr. Christer Wingren
Dept. of Immunotechnology
Lund University
BMC D13
SE-22184 Lund
Sweden
+46-46-2224323

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