Principle of Olink Proteomics

Olink proteomics is an advanced technology based on Proximity Extension Assay (PEA), allowing the simultaneous measurement of multiple target proteins with exceptional sensitivity and specificity. The core concept of PEA involves the use of antibodies that specifically bind to target proteins, coupled with precise protein quantification using DNA sequence tags attached to the antibodies.

 

Olink's PEA technology merges antigen-antibody interactions with DNA extension techniques. Specifically, Olink employs a pair of antibodies that bind to distinct epitopes on the target protein. These antibodies are conjugated to complementary oligonucleotide sequences. When the target protein is present, the two antibodies come into close proximity, enabling the oligonucleotides to extend and form a double-stranded DNA fragment. This DNA fragment is then amplified using quantitative PCR or similar amplification techniques to quantify protein levels.

 

Antibody Specificity

Olink ensures highly accurate protein detection by using meticulously selected antibodies with strong specificity for their target proteins. These antibodies undergo rigorous validation to confirm their ability to bind specifically to the target protein, minimizing non-specific interactions that could lead to background noise.

 

Signal Amplification

A major advantage of Olink technology is its signal amplification system, which differs from conventional immunoassays like ELISA. Instead of amplifying the protein signal directly, Olink amplifies DNA sequences, thereby increasing detection sensitivity. This approach allows for the reliable detection of proteins at very low concentrations, making it suitable for analyzing a variety of clinical samples and complex biological matrices.

 

Multiplex Detection Capability

Olink's proteomics platform excels at multiplexing, meaning it can measure hundreds of protein expression levels simultaneously in a single experiment. This capability is vital for studying complex biological systems and interactions. Multiplexing is achieved through the high specificity of antibodies, with each antibody pair labeled with distinct DNA tags that allow clear differentiation between signals.

 

Data Processing and Quantification

Once signal amplification is complete, the data is processed by measuring the signal intensity using quantitative PCR equipment. The DNA signal is correlated with the target protein concentration through specialized algorithms. Olink employs normalization strategies, incorporating both internal and external standards, to ensure that the quantification is accurate and consistent.