Surrogate antigens as targets for proteome-wide binder selection

Area of Science:

• Biotechnology
• Immunology
• Proteomics

Background:

• Developing antibodies for proteome-wide studies and biomarker validation is crucial.
• Phage display offers advantages over traditional antibody generation methods like hybridoma technology.
• A key challenge is the limited availability of suitable antigens for proteome-wide antibody selection.

Purpose of the Study:

• To demonstrate the feasibility of using protein epitope signature tags (PrESTs) as surrogate antigens.
• To retrieve target-specific antibody binders using PrESTs in phage display selections.
• To validate the functionality of selected antibody binders for various applications.

Main Methods:

• Utilized cancer-associated PrESTs as surrogate antigens for phage display selections.
• Performed phage selections against PrESTs to identify target-specific binders.
• Validated the selected binders using western blot, immunohistochemistry, and protein microarray.

Main Results:

• Successfully demonstrated the proof of principle for using PrESTs as surrogate antigens.
• Retrieved target-specific antibody binders against various protein targets.
• Confirmed the functionality of these binders in downstream applications like western blot and immunohistochemistry.

Conclusions:

• PrESTs are a viable and accessible alternative to full-length proteins for antibody selection.
• This method facilitates the development of affinity reagents for proteome-wide studies and biomarker discovery.
• The validated binders can be applied in various research and diagnostic applications.