Author: Karen N. Barnard; Brynn K. Alford-Lawrence; David W. Buchholz; Brian R. Wasik; Justin R. LaClair; Hai Yu; Rebekah Honce; Stefan Ruhl; Petar Pajic; Erin K. Daugherity; Xi Chen; Stacey L. Schultz-Cherry; Hector C. Aguilar; Ajit Varki; Colin R. Parrish
Title: The effects of modified sialic acids on mucus and erythrocytes on influenza A virus HA and NA functions Document date: 2019_10_10
ID: mgdqixt0_19
Snippet: These NA VLPs were first tested for enzymatic activity using a standard NA cleavage assay using methylumbelliferyl N-acetylneuraminide (MuNANA) as the substrate (Fig. 5C) (42) . The NA-expressing VLPs were incubated with BSM or with RBCs, and the released Sia were collected and analyzed by HPLC. For BSM, HPLC profiles of total Sia were created to compare NA cleavage preferences (Fig. 6A) . These profiles showed the Sia forms that were susceptible.....
Document: These NA VLPs were first tested for enzymatic activity using a standard NA cleavage assay using methylumbelliferyl N-acetylneuraminide (MuNANA) as the substrate (Fig. 5C) (42) . The NA-expressing VLPs were incubated with BSM or with RBCs, and the released Sia were collected and analyzed by HPLC. For BSM, HPLC profiles of total Sia were created to compare NA cleavage preferences (Fig. 6A) . These profiles showed the Sia forms that were susceptible 255 to NA cleavage and release while the non-released forms were considered to be resistant to NA. These HPLC profiles were then compared to the total Sia released chemically by acid hydrolysis, an unbiased method that removes all Sia forms present in the original sample (45) .
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