Development of proximity probing assays for sensitive protein analysis

UPTEC X 11 001

Examensarbete 30 hp Januari 2011

Development of proximity probing assays for sensitive protein analysis

Andreas Smedjebacka

Molecular Biotechnology Programme

Uppsala University School of Engineering

UPTEC X 11 001 Date of issue 2011-01 Author

Andreas Smedjebacka

Title (English)

Development of proximity probing assays for sensitive protein analysis

Title (Swedish) Abstract

While precise and sensitive detection of proteins is essential to both research and diagnostics, the basic technologies incorporated in the immunoassays of today may be the foundation of tomorrow’s diagnostic tests for early stage cancer. Developed by Olink Bioscience, the proximity extension assay is an in solution immunoassay intended to be applied to biological fluid samples such as blood plasma. By the use of proximity probes carrying DNA oligomers, the assay combines the specificity and variability of antibody-antigen interactions with the sensitivity of real-time PCR detection of nucleic acid strands. This work investigates the formulation of the real-time PCR signal detection step and scouts the underlying principles of unspecific probe-probe interactions. Results recommend parting signal generation and

amplification steps into separate reactions and a composition for the latter is proposed. Also, future approaches to prevent unspecific probe-probe interactions are suggested.

Keywords

Protein detection, immunoassay, in solution, blood plasma, proximity probes, real-time PCR, molecular beacon, DNA polymerase, unspecific probe-probe interactions

Supervisors

Simon Fredriksson Olink AB Scientific reviewer

Mats Nilsson Uppsala University

Project name Sponsors

Language

English ^Security

ISSN 1401-2138 Classification

Supplementary bibliographical information Pages

43

Biology Education Centre Biomedical Center Husargatan 3 Uppsala Box 592 S-75124 Uppsala Tel +46 (0)18 4710000 Fax +46 (0)18 471 4687

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