Page 38 - Pharmaceutical- Guide to Biopharmaceutical

Characterization Quality Control

Measuring Protein Aggregates (with temperature control and mixing) Aggregates Sizer

Accelerated Testing of Protein Stability Using the benefits

Aggregates Sizer TC (With Temperature Control) Cell Line Optimization
click here • With a single system, it is possible to measure all aggregates within the 0.1 to 10 μm
range at the same time.
Operating Principle and Features Results
• Real-time measurements can be performed while controlling the temperature and
The Aggregates Sizer aggregation analysis system for biopharmaceuticals A comparison of aggregate formation during the 40 minutes at 23 °C applying stirring stress.
can measure all aggregates within the previously difficult-to-measure for the 0.2 to 2 μm range versus the 2 to 10 μm range is shown in Fig.
0.1 to 10 μm range at the same time, measure aggregates while 2. It indicates that aggregate formation was greatest in the 2 to 10 • Micro quantities (125 μL) can be measured using a micro cell. Culture
applying a stress, and perform quantitative measurements. The μm range using the PEEK stirrer, and greatest in the 0.2 to 2 μm range
Aggregates Sizer is a particle size analyzer that measures particle using the stainless steel stirrer. The glass stirrer resulted in the least
diameters based on the laser diffraction method. Normally, using the aggregate formation in both ranges. The time-course changes in the
laser diffraction method, particle size can be determined based on a particle size distribution for the PEEK stirrer at 42 °C (Fig. 3) show that
sample's scattering patterns but concentration cannot be determined. aggregate formation occurs as a function of time. It also confirms that
However, the Aggregates Sizer can measure absolute concentration aggregate formation depends on temperature.
values by calibrating with a standard sample of known particle sizes
and concentrations. Quantity of 0.2 to 2 μm Aggregates Formed
q (count/mL)
Measurement Method and Conditions Purification
4.5E+08
The sample solution was prepared by mixing freeze-dried bovine
gamma globulin with PBS (pH 7.4) to a concentration of 1 mg/mL. 5 Concentration (cumulative) 6.0E+08
3.0E+08
mL of the sample solution was measured in a temperature-controlled
batch cell (Fig. 1) while stirring for 40 minutes at 190 strokes per 1.5E+08
minute. Stirring rods made of three materials, PEEK, stainless steel, and 0.0E+00
glass, were used for comparison. A temperature-controlled circulator PEEK SUS GLASS
was used during accelerated testing to maintain the temperature at
three different constant temperature levels (23, 30, and 42 °C) for Quantity of 2 to 10 μm Aggregates Formed
comparison. 4.0E+04 q (count/mL) Characterization
Concentration (cumulative) 3.0E+04

2.0E+04

1.0E+04
0.0E+00 Specifications
PEEK SUS GLASS
Fig. 2 Comparison of Aggregate Formation in Each Size Range
Fig. 1 Batch Cell (with temperature control function) Using Different Stirring Rod Materials (after 40 minutes at 23 °C) Instrument Aggregates Sizer
q (count/mL) Measurement principle *1 Laser diffraction method Quality Control
1e+07 1e+07
Measurement range Particle size range: 100 nm to 10 µm
Initial (Size distribution displayed: 40 nm to 20 µm)
8e+06 10 min 8e+06 Concentration measurement accuracy *2 ± 30 or less
20 min
Concentration (frequency) 6e+06 6e+06 Concentration range *3 Particle size 100 nm: 2 µg/mL to 12 µg/mL
40 min
Particle size 1 µm: 0.5 µg/mL to 10 µg/mL
Particle size 10 µm: 10 µg/mL to 180 µg/mL
4e+06
4e+06
Required liquid volume: 5 mL
2e+06 2e+06 Batch cell Cell material: Quartz glass Pharmacokinetics
Stirring mechanism: Up-and-down movement of blade
Stirring plate material: Stainless steel, glass, PEEK
0 0 Temperature range with temperature control function: 20 to 42 °C ± 1 °C, set from PC
0.05 0.1 0.5 1 5 10 Operating environment: Temperature: 10 to 30 °C Humidity: 20 to 80 % (with no condensation)
Particle Size (µm)
Fig. 3 Aggregate Formation Process during Accelerated Testing (with PEEK at 42 °C) Micro cell Material: Quartz glass and PTFE (cell cap with temperature control function)
Required liquid volume: Approx. 0.125 mL
Conclusion Application Examples (Shimadzu Application News No.) Temperature range with temperature control function: 20 to 42 °C ± 1 °C, set from PC
Operating environment: Temperature: 10 to 30 °C Humidity: 20 to 80 % (with no condensation) Others
The Aggregates Sizer is ideal for evaluating protein stability, because • Evaluating the stability and responsiveness to stress of biopharmaceuticals Dimensions and weight Approx. W 680 mm × D 280 mm × H 430 mm, and approx. 31 kg
it is able to measure aggregate concentrations in real time as stirring • Considering additives for inhibiting aggregation (Q117)
stress is applied under temperature-controlled conditions and as the • Evaluating the concentration of particles in blood (Q120) *1 The measurement range depends on particle characteristics, such as shape.
*2 Concentration measurement accuracy values were measured using reference samples and procedures specified by Shimadzu.
materials that contact liquid are varied. *3 The concentration range depends on particle characteristics, such as shape.

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