Antibody-Linked Fluorogen-Activating Proteins for Antigen Detection and Cell Ablation

Area of Science:

• Biochemistry
• Molecular Biology
• Cell Biology

Background:

• Selective labeling of cell surface proteins is crucial for biological research and therapeutic interventions.
• Existing methods may have limitations in specificity or applicability.

Purpose of the Study:

• To develop a versatile method for selective labeling of cell surface proteins using fluorogen-activating proteins (FAPs) conjugated to immunoglobulins (IgGs).
• To demonstrate the utility of FAP-antibody conjugates in various biological applications.

Main Methods:

• Conjugation of FAPs to standard IgGs using a novel polypeptide reagent.
• The reagent features a photoactivatable Fc-binding domain and a FAP domain.
• Application of FAP-antibody conjugates in cultured mammalian cells.

Main Results:

• Successful selective labeling of cell surface proteins was achieved.
• FAP-antibody conjugates demonstrated efficacy in protein detection and cell ablation.
• Visualization of cell-cell contacts was enabled using a tethered fluorogen assay.

Conclusions:

• The developed FAP-antibody conjugation strategy offers broad utility for experimental and therapeutic applications.
• This approach is adaptable to most commercially available IgGs.
• The method provides a powerful tool for cell surface protein analysis and manipulation.