School of Biotechnology
Proteomics
Achievements

  Biosensor analysis of DNA manipulations
Research groups
Summary
Human protein atlas
Mycoplasma mycoides
Antibody proteomics
Alkali stabilization
Tree genomics
Pyrosequencing
Single cell analysis
Affibodies
Biosensor (DNA)
Bioautomation
Surface display
Protein folding
In vivo stabilization
Solid phase methods
Protein G
Protein A
Affinity tags



A surface plasmon resonance based optical device for mass sensitive detection of biomolecular interactions at a chip surface was utilized for the first time for quantitative real-time analysis of the action of DNA manipulating enzymes, such as ligase, restriction nucleases and polymerases (1). Sequence-based DNA analysis, first introduced with oligonucleotide model systems, was extended to the scanning and screening for mutations in PCR amplified DNA from clinically relevant samples (2,3). The experimental set-up for mutation detection was further developed by the introduction of a subtractive mismatch sensitive hybridization outside the instrument and a subsequent determination of the relative amounts of remaining oligonucleotides with analytical biosensor monitoring of hybridizations between fully complementary oligonucleotides (4,5). In conclusion, the applied technology was found to be a suitable tool for a wide range of molecular biology applications, with emphasis on hybridization analysis and mutation detection.

Key (own) publications:
1.
Nilsson, P., Persson, B., Uhlén, M. and Nygren, P-Å., (1995). Real-time monitoring of DNA manipulations using biosensor technology. Anal. Biochem. 224: 400-408.
2.
Nilsson, P., Persson, B., Larsson, A., Uhlén, M. and Nygren, P-Å., (1997). Detection of mutations in PCR products from clinical samples by surface plasmon resonance. J. Mol. Recognit. 10:7-17.
3.
Persson, B., Stenhag, K., Nilsson, P., Larsson, A., Uhlén, M. and Nygren, P-Å., (1997). Analysis of oligonucleotide probe affinities using surface plasmon resonance: a means for mutational scanning. Anal. Biochem. 246: 34-44.
4.
Nilsson, P., Larsson, A., Lundeberg, J., Uhlén, M. and Nygren, P-Å., (1999). Mutational scanning of PCR products by subtractive oligonucleotide hybridization analysis. BioTechniques, 26:308-316.
5.
Nilsson, P., O’Meara, D., Edebratt, F., Persson, B., Uhlén, M., Lundeberg, J. and Nygren, P-Å., (1999). Quantitative investigation of the modular primer effect for DNA and peptide nucleic acid hexamers. Anal. Biochem., 269: 155-161.
6.
O´Meara D., Nilsson P., Nygren P-Å, Uhlén M. and Lundeberg J. Capture of single stranded DNA assisted by oligonucleotides modules. 1998, Anal Biochem 255, 195-203
Last updated: 2010-12-22