Reversible interference of Fe³(+) with monoclonal antibody analysis in cation exchange columns.
The effect of Fe³(+) treatment on weak cation exchange column chromatography was demonstrated for monoclonal antibody (MAb) analysis. Fe³(+)-exposed columns showed lowered relative peak areas of the parent MAb peak as well as both acidic and basic variant peaks that could lead to erroneous conclusions. Accurate measurement of relative amounts of variants to the parent MAb is essential for demonstrating the safety and efficacy of therapeutic molecules such as MAbs. Complete reversal of the compromised MAb analysis performance was observed after washing the column with chelating agents, confirming that metal contamination was responsible for the compromised column performance.
R&D, Dionex Corporation, Sunnyvale, CA 94085, United States. srinivasa.rao@dionex.com
Srinivasa
S
Rao
Chris
C
Pohl
D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000737:chemistry; D002412:Cations / Q000737:chemistry; D002614:Chelating Agents / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods*; D005290:Ferric Compounds / Q000737:chemistry*; D053758:Nanoparticles / Q000737:chemistry
D000911:Antibodies, Monoclonal; D002412:Cations; D002614:Chelating Agents; D005290:Ferric Compounds