Antibodies labeled with small molecules such as fluorophore, biotin or drugs play a critical role in various areas of biological research,drug discovery and diagnostics. There are several limitations to current methods for labeling antibodies including the need for purified antibodies at high concentrations and multiple buffer exchange steps.
In a recent publication, a method (on-bead conjugation) is described that addresses these limitations by combining antibody purification and conjugation in a single workflow. This method uses high capacity-magnetic Protein A or Protein G beads to capture antibodies directly from cell media followed by conjugation with small molecules and elution of conjugated antibodies from the beads.
Using a variety of fluorophores the researchers show that the on-bead conjugation method is compatible with both thiol- and amine-based chemistry.
This method enables simple and rapid processing of multiple samples in parallel with high-efficiency antibody recovery. It is further shown that recovered antibodies are functional and compatible with downstream applications.
Nidhi, N. et al. (2015) On-bead antibody-small molecule conjugation using high-capacity magnetic bead J. Immunol. Methods http://dx.doi.org/10.1016/j.jim.2015.08.008
Latest posts by Gary Kobs (see all)
- Optimizing Pressure Cycling Sample Preparation for Bottom-Up Proteomics - February 11, 2019
- Understanding Mechanisms of Pesticide Resistance to Thiamethoxam in the Cotton Aphid - January 11, 2019
- Cell Free Application: Characterization of Long Non-coding RNA Inhibition of Transcription - November 12, 2018