[RASMB] Characterizing hetero association via sedimentation equilibrium

[Toth IV, Ronald T]

Hello Members!

Thank you all for your help in the past! I have a new question!

Given that the standard procedure for characterizing a self association with sedimentation equilibrium is to sediment three different concentrations at three different rotor speeds, my question is this: Is there a standard procedure for characterizing a hetero association?

Should I sediment three samples at different total protein concentration while keeping the molar ratio of antibody to ligand constant? Should I sediment three different molar ratios of antibody to ligand?

Some further information: We have an antibody-ligand system, and the ligand is 20 kDa. The ligand binds in the Fc region, so it's not necessarily limited to a divalent interaction.

Thanks again for your help I really appreciate it!

- Ronald Toth

Post Doctoral Researcher
Macromolecule and Vaccine Stabilization Center
University of Kansas
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