Purification and Characterization of FliC and CRM197
STm phase 1 flagellin proteins (FliC) were purified as described from culture supernatants of CVD 1925 [20 (link)]. Recombinant CRM197 produced in E. coli was obtained from Fina Biosolutions (Rockville, MD). FliC and CRM197 were confirmed for integrity and removal of residual host cell protein by SDS-PAGE with Coomassie Brilliant Blue staining, and endotoxin levels by Limulus Amebocyte Lysate (LAL) assay using the Endosafe PTS system (Charles River Laboratories, MA).
Baliban S.M., Yang M., Ramachandran G., Curtis B., Shridhar S., Laufer R.S., Wang J.Y., Van Druff J., Higginson E.E., Hegerle N., Varney K.M., Galen J.E., Tennant S.M., Lees A., MacKerell AD J.r., Levine M.M, & Simon R. (2017). Development of a glycoconjugate vaccine to prevent invasive Salmonella Typhimurium infections in sub-Saharan Africa. PLoS Neglected Tropical Diseases, 11(4), e0005493.
Removal of residual host cell protein in FliC and CRM197
Endotoxin levels in FliC and CRM197
control variables
Recombinant CRM197 produced in E. coli
controls
Positive control: Recombinant CRM197 produced in E. coli
Negative control: Not specified
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