Separation of human immunoglobulin G subclasses using monolith technology
Conference Paper
Overview
Overview
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.
