Briefly, polySia avDP20 was oxidized to an aldehyde by sodium metaperiodate, and then hydrazide coupled biotin (Thermo Scientific) was conjugated to form a hydrazone bond. AAV6 and newly identified type 6-like variants also required...
HepG2 (A) and Pro-5 (B) cells were transduced with a constant amount of AAV1-luc or AAV6-luc vectors in the presence of a 200-fold-excess amount of competing AAV1, AAV6, or AAV2 encapsidated λ phage DNA-containing...
The data from these assays were complemented by a glycan array analysis which determined that AAV1 efficiently binds to NeuAcα2-3GalNAcβ1-4GlcNAc as well as glycoproteins containing NeuAcα2-3/α2-6Galβ1-4GlcNAc N-linked glycans. Sialic acids are typically found as...
In some experiments the bacterial surface was desialylated with 13· In the earlier set of experiments that utilized neuraminidase, we noted that, following neuraminidase treatment, there were typically fewer cells in the dish, indicating...
To assay for oxidoreductase activity, the purified recombinant proteins were incubated in 100-μl reactions at 37 °C overnight with 1 mg/ml 2,7-anhydro-Neu5Ac or Neu5Ac in 20 mm sodium phosphate buffer, pH 7.5, in the...
We stained cell lines with lectins that have been used to recognize the following three different epitopes: (i) WGA recognizes all sialic acids, (ii) MAA recognizes α2,3 sialic acid, and (iii) SNA recognizes α2,6...
