Influence of antibody-antigen interaction on fluorescence of Rhodamine 6G conjugated to the antibodies measured in human serum
Fluorescent dyes conjugated to monoclonal antibodies specifically trastuzumab (TRAS) and pertuzumab (PERT) serve as sensitive reporters of changes in the local energetic environment arising from antibody-antibody and antibody-antigen interactions. Upon covalent conjugation, the antibody local environment perturbates the energetic state of the dye, resulting in changes of the fluorescence emission…
Monoclonal antibody-conjugated fluorescent dyes, specifically trastuzumab (TRAS) and pertuzumab (PERT), can detect changes in the local energetic environment arising from antibody-antibody and antibody-antigen interactions. Covalent attachment of these antibodies alters the dye's fluorescence emission energy and emission width. Researchers measured these spectral parameters in undiluted and diluted human serum, as well as in a human serum albumin (HBS-8) solution.
They discovered that antibody-antigen interactions significantly impact emission energy and width, which vary with the surrounding medium's composition. The study found that TRAS exhibited the most pronounced spectral shifts at 100 nM concentration in serum when interacting with the HER2 extracellular domain. This research suggests that analyzing fluorescence emission energy and width can be an effective method for probing antibody interactions in complex biological matrices under conditions similar to those found in the human body.
Written by urgent.news from bioRxiv's reporting — not their text. Machine-written — may contain errors; check the original before relying on it.
