a carboxy-terminal trimerization domain stabilizes conformational epitopes on the stalk domain of soluble recombinant hemagglutinin substrates一个carboxy-terminal三聚域杆的稳定构象表位域的可溶性重组血凝素基质.pdfVIP

A Carboxy-Terminal Trimerization Domain Stabilizes Conformational Epitopes on the Stalk Domain of Soluble Recombinant Hemagglutinin Substrates 1 1,3 1 1,3 1 1,2 Florian Krammer , Irina Margine , Gene S. Tan , Natalie Pica , Jens C. Krause , Peter Palese * 1 Department of Microbiology, Mount Sinai School of Medicine, New York, New York, United States of America, 2 Department of Medicine, Mount Sinai School of Medicine, New York, New York, United States of America, 3 Graduate School of Biological Sciences, Mount Sinai School of Medicine, New York, New York, United States of America Abstract Recently, a new class of broadly neutralizing anti-influenza virus antibodies that target the stalk domain of the viral hemagglutinin was discovered. As such, induction, isolation, characterization, and quantification of these novel antibodies has become an area of intense research and great interest. Since most of these antibodies bind to conformational epitopes, the structural integrity of hemagglutinin substrates for the detection and quantification of these antibodies is of high importance. Here we evaluate the binding of these antibodies to soluble, secreted hemagglutinins with or without a carboxy-terminal trimerization domain based on the natural trimerization domain of T4 phage fibritin. The lack of such a domain completely abolishes binding to group 1 hemagglutinins and also affects binding to group 2 hemagglutinins. Additionally, the presence of a trimerization domain positively influences soluble hemagglutinin stability during expression and purification. Our findings suggest that a carboxy-terminal trim