1. test your passed through solvent when loading samples to make sure that no sample is trapped in the column. 2. Use another batch of antibody. 3. make sure that your sample doesn't denature. 4. check the solvent you used to prepare sample solution. 5. check if you used the right resin. 

Hi there,

Your elution buffer is not actually a buffer as pH 2.4 (by the way I have in mind that 0.1M acetic acid gives a pH around 2.9) is far from acetic acid pKa (which is around 4.8).Therefore the pH may be affected when in contact of the column matrix. For optimizing elution please read the following from ThermoFisher:

https://tools.thermofisher.com/content/sfs/brochures/TR0027-Elution-conditions.pdf

Ok, there is A way out of it. Wash the column thoroughly with the elution buffer so that no protein is detected in the void volume by simply checking it at 280nm. Then start the experiments all over again. The problem should disappear.