title: Comprehensive analysis of pathogen-specific antibody response in vivo based on an antigen library displayed on surface of yeast.
abstract: Host antibody response is a crucial defense against pathogenic infection. Here, we report a novel technique allowing quantitative measurement of polyclonal antibody response in vivo. This involves expression of a combinatorial library of target proteins from a candidate pathogen on the surface of yeast Saccharomyces cerevisiae. After mixing with serum/plasma from infected or immunized subjects, positive yeast clones were isolated via fluorescence-activated cell sorting (FACS). Using this technique, we have studied mouse immunized serum with recombinant hemagglutinin (HA) protein from a human influenza H5N1 strain (A/Anhui/1/2005) and convalescent plasma from an infected human in China. Our technique has identified novel antigenic domains targeted by serum/plasma and allowed calculation of the relative proportion of the antibody response against each domain. We believe such systematic measurement of an antibody response is unprecedented, and applying this method to different pathogens will improve understanding of protective immunity and guide development of vaccines and therapeutics.
authors.affiliation: Comprehensive AIDS Research Center, School of Medicine, Tsinghua University, Beijing 100084, China.
authors.forename: Teng
authors.initials: T
authors.lastname: Zuo
authors.forename: Xuanling
authors.initials: X
authors.lastname: Shi
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Liu
authors.forename: Linlin
authors.initials: L
authors.lastname: Guo
authors.forename: Qing
authors.initials: Q
authors.lastname: Zhao
authors.forename: Tianxia
authors.initials: T
authors.lastname: Guan
authors.forename: Xianming
authors.initials: X
authors.lastname: Pan
authors.forename: Na
authors.initials: N
authors.lastname: Jia
authors.forename: Wuchun
authors.initials: W
authors.lastname: Cao
authors.forename: Boping
authors.initials: B
authors.lastname: Zhou
authors.forename: Mark
authors.initials: M
authors.lastname: Goldin
authors.forename: Linqi
authors.initials: L
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000917:Antibody Formation / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D000956:Antigens, Viral / Q000737:chemistry / Q000276:immunology*; D002681:China; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000737:chemistry / Q000276:immunology; D006801:Humans; D007114:Immunization; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology*; D007251:Influenza, Human / Q000276:immunology / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009976:Orthomyxoviridae Infections / Q000276:immunology / Q000821:virology; D019151:Peptide Library; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000276:immunology*; D013045:Species Specificity
chemical_list: D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D019151:Peptide Library