Page 6 - Pharmaceutical M5 Biopharma

6  Sponsored Feature

Intact Proteins Robust and reliable separation and measurement

Aggregate and Fragment Analysis

Purified HPLC (SEC)
Protein

Protein aggregates are known to strongly
induce immunogenic responses when
administered, and aggregation is one of
the most important quality attributes
that must be monitored and managed
at every step of process development
and refinement. For example, in the initial
stage of development, cell lines that have
greater tendency to produce aggregates
or degradants are screened out even if it
showed a high secretion yield.
Size-exclusion chromatography (SEC)
is the method of choice for separating
the major monomer fraction from size
variants occurring by either aggregation
or degradation. Shimadzu supplies the
range of SEC columns that are useful
particularly in increasing the throughput
needed for the screening analysis.

Size-exclusion chromatography of mAb
Column: Shim-pack Bio Diol-300
Flow rate: 0.2 mL/min
Sample: Humanized monoclonal IgG1

Here, the chromatograms compare
the SEC separation profile of mAb
acquired by Shim-pack Bio Diol-300
columns of different dimension at a
constant flow rate. Reduction of particle Column N (3) Rs (1,2) Rs (2,3)
size (5 μm to 2 μm) resulted in increased (1) 5 μm, 300 x 4.6 mm 8,500 0.88 2.67
peak resolution, which in turn gave the (2) 2 μm, 300 x 4.6 mm 16,200 1.17 4.15
room to reduce the analysis time by (3) 2 μm, 150 x 4.6 mm 8,700 0.85 2.75
using a shorter column (30 minutes to
15 minutes).