Sequentially-Activated Antibodies Based on Programmable DNA Tags for Rapid Multiplexed Protein Imaging

Area of Science:

• Biotechnology
• Molecular Imaging
• Immunofluorescence Assays

Background:

• Multiplexed protein imaging is crucial for biological research but hindered by lengthy protocols and low signal.
• Existing methods often require extensive washing steps and stimulating reagents, limiting speed and efficiency.

Purpose of the Study:

• To develop a novel method for rapid, wash-free sequential imaging of multiple protein targets.
• To overcome the limitations of current multiplexed protein imaging techniques.

Main Methods:

• Development of sequentially-activated antibodies (SAAs) by conjugating DNA tags to primary antibodies with signal enhancement.
• Simultaneous pre-processing of samples to label all targets with SAAs.
• Wash-free sequential signal activation and detection using strand displacement reactions with rapid kinetics (t_1/2 < 10 s).

Main Results:

• Demonstrated rapid, wash-free sequential imaging of nine different protein targets.
• Achieved imaging within minutes, significantly reducing assay time.
• Showcased high signal intensity and efficient target detection.

Conclusions:

• Sequentially-activated antibodies (SAAs) provide an efficient all-in-one platform for rapid multiplexed protein imaging.
• This method offers a promising solution for various applications in immunofluorescence imaging.
• The wash-free and rapid nature of SAAs accelerates biological discovery.