Page 25 - Shimadzu Journal vol.9 Issue2

Biopharmaceutical

High Performance Liquid Chromatograph

Prominence™ Inert HPLC System, LH-40

Seamless Process from

Protein Purification to Evaluation

Risa Suzuki 1
1 Global Application Development Center, Analytical & Measuring Instruments Division, Shimadzu Corporation

User Benefits
・ By choosing target peaks after fractionation, appropriate fractions can Seamless Analysis with Liquid Handler
be automatically reinjected for analysis.
・ Equipped column­switching valve allows automatic column switching The liquid handler (LH-40) serves as an autosampler as well as a
between purification and analysis.
fraction collector for the LC system. That means samples fraction-
・ It is also convenient for comparing several samples to optimize
culture conditions. ated during the first run can be injected directly for the second
run without transferring them from a fraction collector to an au-
tosampler. For example, with this system (Fig. 1 and 2), the target
Introduction protein is purified by an affinity column and fractionated as the
first step, and then the fractionated protein elution is re-injected for
Size exclusion chromatography (SEC) is one technique used to SEC analysis as the second step. These two steps can be performed
separate molecules based on size, and it is one of the main meth- simply by specifying the method and fraction.
ods used for verification of protein multimer formation. However, In this example, we evaluated IgG in human plasma using an
this technique can not distinguish proteins from ones with simi- LH-40 liquid handler installed in a Prominence inert LC system.
lar molecular sizes. Because smaller molecules can enter pores of
materials packed in an SEC column, but larger molecules cannot.
Therefore, larger molecules elute earlier than smaller molecules. In
the case of target proteins in serum or culture supernatant, the sam-
ples have to be purified before SEC analysis. This article describes
a seamless process of target proteins for purification, fractionation,
and re-injection utilizing a liquid handler.

Shimadzu Journal vol.9 Issue2 58