Comprehensive Comparison of AAV Purification Methods: Iodixanol Gradient Centrifugation vs. Immuno-Affinity Chromatography

<table class="figure-group"><tr class="fig-image" id="a"><td><object data="https://static.hindawi.com/articles/acgt/volume-2023/2339702/figures/2339702.fig.002a.svgz" name="2339702.fig.002a" type="image/svg+xml"></object></td></tr><tr class="fig-caption"><td><b>(a) </b>Suspension HEK293 cells</td></tr><tr class="fig-image" id="b"><td><object data="https://static.hindawi.com/articles/acgt/volume-2023/2339702/figures/2339702.fig.002b.svgz" name="2339702.fig.002b" type="image/svg+xml"></object></td></tr><tr class="fig-caption"><td><b>(b) </b>Subculture for transfection</td></tr></table>

<div>Viability and viable cell density (VCD) representation of scalable suspension system HEK293 cells: freshly thawed cells (a) and subcultured cells (b). Transfection was performed on day 2 with a VCD of ~2.0–<svg height="11.9413pt" id="M2" style="vertical-align:-0.3499002pt" version="1.1" viewbox="-0.0498162 -11.5914 46.5453 11.9413" width="46.5453pt" xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink"><g transform="matrix(.013,0,0,-0.013,0,0)"><path d="M412 140C382 77 369 73 315 73H129L270 222C362 320 402 379 402 466C402 571 322 635 234 635C177 635 130 609 99 576L42 495L64 475C90 514 133 568 201 568C274 568 318 519 318 435C318 349 255 267 193 193C144 135 87 78 32 23V0H405C417 45 427 89 440 131L412 140Z"></path></g><g transform="matrix(.013,0,0,-0.013,6.24,0)"><path d="M113 -12C146 -12 170 11 170 46C170 78 146 103 114 103S58 78 58 46C58 11 82 -12 113 -12Z"></path></g><g transform="matrix(.013,0,0,-0.013,9.204,0)"><path d="M153 550H386L412 615L406 623H120L82 318C104 327 142 338 184 338C294 338 347 275 347 187C347 112 305 39 221 39C160 39 119 71 97 89C88 97 80 96 71 90C59 80 50 67 49 57C48 45 52 36 66 23C80 9 123 -12 169 -12C221 -11 288 15 342 59C403 109 431 165 431 225C431 308 366 395 238 395C212 395 165 379 127 364L153 550Z"></path></g><g transform="matrix(.013,0,0,-0.013,18.35,0)"><path d="M528 54L331 254L528 455L492 493L294 291L96 493L60 455L257 254L60 54L96 16L294 217L492 16L528 54Z"></path></g><g transform="matrix(.013,0,0,-0.013,28.886,0)"><path d="M384 0V27C293 34 287 42 287 114V635C232 613 172 594 109 583V559L157 557C201 555 205 550 205 499V114C205 42 199 34 109 27V0H384Z"></path></g><g transform="matrix(.013,0,0,-0.013,35.126,0)"><path d="M241 635C89 635 35 457 35 312C35 153 89 -12 240 -12C390 -12 443 166 443 312C443 466 390 635 241 635ZM238 602C329 602 354 454 354 312C354 172 330 22 240 22C152 22 124 173 124 313S148 602 238 602Z"></path></g><g transform="matrix(.0091,0,0,-0.0091,41.413,-5.741)"><path d="M141 347C171 480 264 541 327 570C364 587 403 599 436 605L429 641C389 634 339 622 300 609C191 570 37 458 37 239C37 86 126 -12 245 -12C367 -12 454 89 454 210C454 314 380 397 272 397C252 397 230 390 208 380L141 347ZM231 338C323 338 366 257 366 174C366 108 340 27 263 27C180 27 129 123 129 244C129 268 130 292 135 310C158 323 193 338 231 338Z"></path></g></svg> cells/mL and viability of &gt;92% and vector harvest was at day 5, which was 72 hr posttransfection.</div>

Advances in Cell and Gene Therapy

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Figure 2

Figure 2: Comprehensive Comparison of AAV Purification Methods: Iodixanol Gradient Centrifugation vs. Immuno-Affinity Chromatography 