AAVQC002-004Alkaline Gel ElectrophoresisAgarose gels6 Capsid characterization, purity and aggregationAAVQC003-001AAV Empty Capsid Rate <AUC>AUC to analyze empty capsid rate5 AAVQC003-002TEM (5 pictures) +reportTEM3-5 AAVQC003-003DLS (Dynamic light scattering)Dynamic light scattering5 ...
iCELLis is applicable to perfusion-cultured HEK293 cells, another mainstay of both AV and AAV vector processing. In HEK293 cells, iCELLis improves cellular productivity by optimizing media and cell attachment, but these modifications, Nieminen says, may require careful optimization to avoid potential ...
After packaging the vectors, alkaline gel electrophoresis and Southern analysis were carried out on purified vector genome DNAs to assess their size (Figure 3b). The results were similar for vectors packaged in AAV2 and AAV5 capsids. Both the 4.7 and 5.2 kb vectors showed clear bands at ...
four backbones was packaged into an AAV8 capsid and the viral DNA was isolated67. Size comparisons using alkaline agarose gel electrophoresis analysis revealed that depending on the sgRNA-121 cassette orientation and position, the genomes of certain dual-sgRNA AAV vectors indeed produced multiple trun...
AAV9 genomes derived from either Sf9 or ExpiSf9 cells were run on an alkaline gel, as previously described.26 Briefly, 1% agarose gel was made in alkaline electrophoresis buffer, which is composed of 50 mM NaOH and 1 mM EDTA. AAV DNA samples were prepared using 6× alkaline sample loadin...
The plasma alkaline phosphatase (ALP) level was higher in homozygous Pigob/b mice than in their wild-type controls (p < 0.05). Pigob/− mice showed even more elevated levels of plasma ALP than their wild-type controls (p < 0.001) (Fig. 1g). As hyperphosphatasia in IGD wit...
The AAV vectors purified with either iodixanol or CsCl gradients were titered using quantitative real-time PCR (qPCR). For this purpose, the DNA was extracted from the AAV vectors using an alkaline lysis protocol, in which 10 µl of the AAV vector sample were mixed with 10 µl TE...
DNA sequences encoding β-lactamase, β-galactosidase (LacZ), alkaline phosphatase, thymidine kinase, green fluorescent protein (GFP), chloramphenicol acetyltransferase (CAT), luciferase, and others well known in the art. When associated with regulatory elements which drive their expression, the ...
(B) Alkaline gel of viral DNA from AAV8GFP_P5NV and AAV8GFP_P5F8 virions (1 kb DNA ladder). (C) Graph of GFP expressing HEK293T cells 72 h after transduction with AAV8GFP_P5NV (purple), AAV8GFP_P5F8 (red), AAV8RevGFP_P5F8 (orange), or AAV8Empty_P5F8 (blue) (one-way ...
AP, alkaline phosphatase. N, No AAV. N/A, non-applicable. wks., weeks post-injection. Data are mean ± SEM. Statistical analysis was performed using Crawford-Howell test for h, i, o One-way ANOVA with Tukey’s multiple comparisons for l, m, and r, and Student’s t-test for ...