Parallel barcoding of antibodies for DNA-assisted proteomics.
Mahya Dezfouli1, Sanja Vickovic, Maria Jesus Iglesias
1Science for Life Laboratory, Division of Gene Technology, School of Biotechnology, Royal Institute of Technology (KTH), Solna, Sweden.
Proteomics
|September 30, 2014
Summary
This study introduces a magnetic bead-based DNA-barcoding method for antibodies, enabling ultra-sensitive proteomics. This approach significantly reduces reagent needs and maintains antibody functionality for multiplexed Immuno-Sequencing applications.
Area of Science:
- Biochemistry
- Molecular Biology
- Proteomics
Background:
- DNA-assisted proteomics offers ultra-sensitive, multiplexed measurements using DNA-barcoded affinity reagents.
- Many available antibodies lack optimal quantity, concentration, or purity for standard bio-conjugation.
Purpose of the Study:
- To develop a novel magnetic bead-assisted DNA-barcoding approach for antibodies.
- To enable efficient and scalable multiplexed proteomics applications.
Main Methods:
- A magnetic bead-assisted DNA-barcoding technique was developed for antibodies.
- The method was validated using sandwich immunoassays and quantitative Immuno-PCR assays.
- Multiplexed antibody DNA-barcoding was demonstrated on suspension bead arrays with sequencing readout (Immuno-Sequencing).
Main Results:
- The DNA-barcoding approach significantly reduced reagent quantities (up to 1000-fold).
- Antibody functionality was retained after DNA-barcoding, as shown in immunoassays.
- Successful application in Immuno-Sequencing for multiplexed proteome analysis was demonstrated.
Conclusions:
- The developed method provides a scalable and efficient way to DNA-barcode antibodies for proteomics.
- This technique facilitates ultra-sensitive multiplexed proteomic analyses, including human plasma sample analysis.
