Anion Exchange Chromatography

 The purpose of the Anion Exchange chromatography is to separate nucleic acid from the
total biomass.
 The majority component of biomass will bound to the resin based to their positively
charged column once a binding buffer is passing through the column and then an elution
buffer is use to separate only successful bind molecules. The binding buffer that will be
utilized is 1.0M NaCl and 50mM MOPS at pH 7.4 and elution buffer composition is
1.25M NaCl and 50mM Tris-HCl at pH 7 4. Thus, the concentration of salt will determine
the binding or eluting properties of plasmid DNA towards resin of Anion Exchange
chromatography column.
 The inlet of Anion Exchange chromatography column is 134.26 kg which consist of total
biomass at 132.26 kg and pDNA at 8.82 kg. The mass of nucleic acid removed at S-118
is 37.04 kg which is 70% from total nucleic acid mass and the remaining biomass is
95.22 kg.
 The ratio of binding buffer to elution buffer is at least 2:3 to ensure the optimal binding
condition. While, the volume of elution buffer is 1 ml for each 4 ml of biomass volume.
Thus 20 kg of binding buffer and 33 kg of elution buffer is needed in the process
Stream No./ Materials IN (kg) OUT (kg)
S-115 -
Biomass 132.26
• Endotoxins 44.1
• Nucleic Acid 52.92
• Small molecules and ions 26.42
• pDNA 8.82
3 M KAc 2
S-116 -
• Binding buffer (1.0M NaCl + 50mM MOPS) 20
S-117 -
• Elution Buffer (1.25M NaCl + 50mM Tris-HCl) 33
S-118 -
3 M KAc 2
Binding Buffer (1.0M NaCl + 50mM MOPS) 20
• Nucleic acid 37.04
S-119 -
Biomass 95.22
• Endotoxins 44.1
• Small molecules and ions 26.42
• Nucleic acids 15.88
• pDNA 8.82
Elution Buffer(1.25M NaCl + 50mM Tris-HCl) 33
TOTAL 187.26 187.26