ProteinSimple eBook - 20

Simplifying Protein Analysis * Comparability Study of Manual and Automated Particle Characterization with MFI

Results
At least eight manual and automated runs were performed at each site, using the same
manual and automated protocols for pipetting and analysis (Figure 1). Data were analyzed
to assess normality of data set, and were not found to signiﬁcantly deviate from normal
distribution. P-values were 0.072 and 0.053 for ProteinSimple and Takeda, respectively.
Variances between automated and manual methods were not statistically different by
f-test. Employing a standard analysis for statistical variance(ANOVA) indicated that only
between 0 and 4% of variance in particle counts can be attributed to method type which is
not signiﬁcant. Regression analysis showed no relationship between other input variables
and variance in the data (Figure 3).
Comparison of data showed results at Takeda were approximately 2000 counts/mL
lower than those at ProteinSimple. This difference is likely related to sample quality in
terms of sample stress or degradation during shipment. Neither site showed any signiﬁcant
difference in counts or concentration between the manual and automated method,
conﬁrming that similar results can be obtained using either format.
Count and concentration for automated versus manual format for 1% BSA,
ProteinSimple. Data from tests performed at ProteinSimple are listed in Table 4.
The automated format of 1% BSA protein solution produced consistent counts and
concentration data, compared to the manual format. The modiﬁcation to Protocol D
reduces the sample ﬂush volume without impacting the consistency of results between
replicates in the automated method. The standard deviation for both methods was 5%
or less for all samples tested.
20

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MANUAL

AUTOMATED

RUN

COUNTS

COUNTS/ML

RUN

COUNTS

COUNTS/ML

8783

11184

8463

10780

9745

12409

8444

10773

8530

10862

8483

10812

8238

10490

9003

11479

Average

8824.00

11236.25

Average

8598.25

10961.00

Std Dev

565.63

720.25

Std Dev

234.09

299.43

%CV

6.41%

%CV

2.72%

2.73%

RUN

COUNTS

COUNTS/ML

RUN

COUNTS

COUNTS/ML

7918

10083

8840

11274

7703

9809

9124

11633

7815

9951

8905

11343

7403

9427

10572

13471

Average

7709.75

9817.50

Average

9360.25

11930.25

Std Dev

192.73

245.40

Std Dev

707.47

899.69

%CV

2.50%

%CV

7.56%

7.54%

AVERAGE OF ALL 8 MEASUREMENTS

Average

8227.83

10477.17

Average

9058.00

11545.67

Std Dev

875.47

1114.73

Std Dev

787.21

1001.27

%CV

10.64%

%CV

8.69%

8.67%

Table 4. Comparison of the Manual Versus Automated Method,
ProteinSimple

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