Sequencing of the influenza PR8 virus hemagglutinin Sb site mutant library

Current flu vaccine induces strong response to plastic and higly variable hemagglutinin (HA) head. However, response to conserved epitopes in HA elicited by current vaccine or infection is very limited. We hypothesized that change the relative "molarity" of immunodominant epitopes and conserved domain might increase response to conserved domains. Therfore, we made a library with the most ommunodominant domian Sb site randomized and tested its efficacy in vivo. Here we show the diversity of the library. Four codons in Sb site of PR8 hemagglutinin were randomized by mutagenesis. Viable mutants were sorted with hemaglutinin stalk trimer monoclonal 6F12 by megnetic-activated cells sort. Library before purification and after purification were sequenced.