The adsorption of clavulanic acid (CA) in a fixed-bed column of layered double hydroxides (LDHME) was investigated. Breakthrough curves were obtained experimentally and the system was evaluated with regards to column operation time, efficiency and productivity as functions of simultaneous variations of superficial velocity (vz) and bed height (L) using a central composite rotatable design (CCRD). At the optimized condition (vz = 1.00 cm/min and L = 6.5 cm), the responses were: 46 min, 146 min, 100 min and 13 kg/h cm3, for breakthrough time (tb), exhaustion time (te), the difference te − tb and productivity (P), respectively. These results represent no change in tb, but a 50% decrease of te, 40% decrease of (te − tb) and 38% increase of P, which is advantageous for the process. The factorial design technique was shown to be an efficient tool for assessing factor influences and was effective in optimizing the column operating conditions. Good separation of CA from the amino acids tyrosine (TYR) and proline (PRO) was observed in fixed bed columns. The LDHME adsorbent was shown to be an excellent alternative for CA purification.