Human Gene Therapy - April 2023 - 331

Figure 3. VG/mL accuracy by recovery using the Wyatt 500A˚ column. (A) Five volume transfers at each level were performed with the process confirmation
material. The mean VG/mL value from Supplementary Table S1 was used to establish the accepted VG/mL value of the sample. The recovery of each injection
at each level is shown with the mean- standard deviation. A dashed line shows -20% of 100% recovery. The complete tabulated data with molar mass
information is shown in Table 1. (B) Representative UV chromatogram from SEC-MALS data in (A). Data from the three highest spike levels are shown. The
cyan, blue, and black traces correspond to the 2.5 · 10,13 1.0 · 10,13 and 2.5 · 1012 VG/mL spike concentrations, respectively. The y-axis is the relative scale from
the UV 260 nm trace. The aggregate peaks are detectable near 6.3 to *7 min elution times. Injection volumes and flow rates were as in Fig. 1. The molar
masses for the DNA, capsid, and total vector symbols are as in Fig. 1.
Table 1. Accuracy by recovery data by size exclusion chromatography with UVand multiangle light scattering
Mean Vector
Spike Concentration
(VG/mL)
1.50E+11
2.50E+11
5.00E+11
1.00E+12
2.50E+12
1.00E+13
2.50E+13
Mean Measured
VG/mL
1.36E+11
2.29E+11
4.91E+11
9.28E+11
2.32E+12
1.04E+13
2.63E+13
%CV
7%
3%
8%
3%
5%
4%
3%
Mean Percent
Recovery
91%
92%
98%
93%
93%
104%
105%
%CV, coefficient of variance; CP, capsid particles; VG, vector genomes.
j 331
Mean DNA
Molar Mass (g/mol) %CV
5.24E+06
5.13E+06
5.16E+06
5.02E+06
5.03E+06
5.00E+06
4.98E+06
5%
1%
4%
1%
1%
0%
0%
Molar Mass (g/mol) %CV
1.28E+06
1.23E+06
1.25E+06
1.22E+06
1.22E+06
1.21E+06
1.20E+06
5%
2%
4%
1%
1%
0%
0%
Mean
VG/CP Ratio
0.950
0.916
0.926
0.927
0.922
0.918
0.916

Human Gene Therapy - April 2023

Table of Contents for the Digital Edition of Human Gene Therapy - April 2023

Contents
Human Gene Therapy - April 2023 - CT1
Human Gene Therapy - April 2023 - CT2
Human Gene Therapy - April 2023 - Cover1
Human Gene Therapy - April 2023 - Cover2
Human Gene Therapy - April 2023 - 239
Human Gene Therapy - April 2023 - 240
Human Gene Therapy - April 2023 - 241
Human Gene Therapy - April 2023 - 242
Human Gene Therapy - April 2023 - 243
Human Gene Therapy - April 2023 - 244
Human Gene Therapy - April 2023 - Contents
Human Gene Therapy - April 2023 - 246
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Human Gene Therapy - April 2023 - Cover3
Human Gene Therapy - April 2023 - Cover4
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