Figure_2.tif (1.1 MB)

Recombinant HA protein expression and purification

Download (0 kB)
posted on 14.06.2013 by Shih-Chang Lin, Jia-Tsrong Jan, Ben Dionne, Michael Butler, Ming-Hsi Huang, Chung-Yi Wu, Chi-Huey Wong, Suh-Chin Wu

. Soluble HA proteins were constructed using the HA cDNA sequences of H5N1 A/Thailand/1(KAN-1)/2004. The PQRERRRKKRG multibasic protease cleavage site between HA1 and HA2 was mutated to PQRETRG to retain the uncleaved protein. The C-terminal sequence was fused with a GCN4pII leucine zipper sequence and ended with a His-tag to facilitate purification. Recombinant H5HA proteins were purified from the culture supernatants of Sf9, Mimic, and CHO cells and analyzed by SDS-PAGE with (A) Coomassie Blue staining and (B) western blotting with anti-H5 antibodies. Proteins were also analyzed following treatment with (C) Endo H and (D) PNGase F.