Analysis of solid-phase immobilized antibodies by atomic force microscopy.

Area of Science:

• Biophysics
• Surface Chemistry
• Analytical Chemistry

Background:

• Antibody adsorption onto solid surfaces can impede molecular interactions.
• Optimizing adsorption conditions is crucial for maintaining antibody functionality.
• Atomic force microscopy (AFM) offers a sensitive, label-free approach for studying molecular interactions.

Purpose of the Study:

• To investigate factors influencing antibody adsorption using AFM.
• To evaluate the efficiency of particle capture by surface-adsorbed antibodies.
• To demonstrate AFM's utility in screening and selecting functional antibodies.

Main Methods:

• Utilized atomic force microscopy (AFM) as a readout mechanism.
• Studied anti-bacteriophage fd antibodies in a solid-phase assay.
• Determined capture efficiencies by varying pH and antibody concentration on gold surfaces.
• Evaluated antibody sensitivity for phage fd capture across different phage concentrations.

Main Results:

• AFM effectively analyzed antibody properties and ligand-antibody interactions.
• Optimized adsorption conditions improved antibody functionality for particle capture.
• Demonstrated the capability to compare capture efficiencies under various surface conditions.
• Showcased the relative sensitivity of antibodies for target capture.

Conclusions:

• AFM is a valuable analytical tool for assessing surface-adsorbed antibody function in capture assays.
• This method facilitates rapid screening and selection of antibodies and ligands.
• The approach is applicable to the growing field of chip-based analytical platforms in proteomics.