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Published on: December 1, 2020
A novel strategy for proteome-wide ligand screening using cross-linked phage matrices
Chen Qian1, Jian-Ning Liu2, Fengyuan Tang1
1Institute of Molecular Medicine and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University Nanjing 210093.
The Journal of Biological Chemistry
|February 2, 2010
Summary
This study introduces a new phage display method for identifying antibody ligands and purifying antigens from complex human plasma proteomes. The approach successfully isolated specific antibody phages and their corresponding target proteins, including low-abundance ones.
Area of Science:
- Biotechnology
- Proteomics
- Immunology
Background:
- Identifying specific ligands within complex biological systems like the human proteome remains a significant challenge.
- Phage-displayed antibody technology offers a powerful platform for molecular recognition and discovery.
Purpose of the Study:
- To develop and validate a novel "library against proteome" panning strategy for simultaneous ligand screening and antigen purification.
- To demonstrate the feasibility of isolating ligands and antigens from a complex, non-comparative proteome-wide system.
Main Methods:
- A solution-phase panning strategy using phage-displayed antibody libraries against the human plasma proteome.
- Biotin/streptavidin beads were utilized for separation over three rounds of panning.
- Selected monoclonal phages were employed as cross-linked matrices for antigen purification.
Main Results:
- Nine monoclonal phages with high affinity for unknown plasma proteins were isolated.
- Six of these phages were successfully used to purify their corresponding antigens.
- Two purified proteins were identified as amyloid protein and apolipoprotein A-I precursor.
Conclusions:
- The developed "library against proteome" panning strategy is effective for discovering antibody ligands and purifying antigens from complex biological samples.
- This method enables the simultaneous isolation of multiple ligands and their targets, including low-abundance proteins.
