Separation of human immunoglobulin G subclasses using monolith technology Conference Paper uri icon

abstract

  • In the present study, a step gradient elution method was evaluated for the separation of human immunoglobulin G (hIgG) into its subclasses on CIM rwprotein A monolithic column. hIgG Seheme 1. Represantative structures of the human IgO subclasses [I). was loaded onto the column and bound protein was eluted with a pH gradient. The subclass content of the eluted fractions was analyzed by enzymewIinked immunosorbent assay (ELISA). Results showed that separation of IgG3 from the other three subclasses can be successfully achieved with high selectivity (100%) and throughput on monolithic media. It was also revealed that enriched fractions of IgG 1 and IgG2 could be obtained from purified hIgG in a 28 minutes long chromatographic run, Three fractions with high IgO 1 content (89.1%,94,3% and 88.8%) were recovered, Furthennore, IgG2 was enriched to 64% successfully. A rapid step gradient elution scheme without any additives in buffers was proven to obtain enriched prep~ions of the two important subclasses with high througbput.

publication date

  • January 1, 2014