Therapeutic monoclonal antibodies (MAb) have rapidly developed into a major class of biological medicines. Registration of growing number of MAbs necessitated need for the establishment of a pharmacopoeial monograph for the minimum quality standards and analytical procedures required for routine quality control of MAb preparations.
Size exclusion chromatography is routinely performed to measure the aggregate level and size variant profiles of MAb preparations. Size exclusion chromatography is one of the test methods included in the draft monograph for MAb analytical procedures proposed by the expert panel of United States Pharmacopoeial convention. The proposed method involves low pressure separation using silica-based size exclusion column (L59) and conventional HPLC systems. Recent advances in Ultra performance liquid Chromatography (UPLC) instrumentation and the development of high pressure size-exclusion columns with sub 2 µm packing made it possible to analyse MAb samples with increased speed and high resolution. Here, we present the results of a preliminary method validation study undertaken in our laboratory to transfer the HPLC-SEC method proposed by USP convention to UPLC-SEC system. Samples of MAb preparations were analysed by HPLC-SEC and UPLC-SEC methods. Purity results obtained by the two methods were statistically analysed to demonstrate that the purity results from UPLC-SEC method are comparable to those obtained by HPLC-SEC method.