title: Identification of antigen-specific human monoclonal antibodies using high-throughput sequencing of the antibody repertoire.
abstract: High-throughput sequencing of the antibody repertoire provides a large number of antibody variable region sequences that can be used to generate human monoclonal antibodies. However, current screening methods for identifying antigen-specific antibodies are inefficient. In the present study, we developed an antibody clone screening strategy based on clone dynamics and relative frequency, and used it to identify antigen-specific human monoclonal antibodies. Enzyme-linked immunosorbent assay showed that at least 52% of putative positive immunoglobulin heavy chains composed antigen-specific antibodies. Combining information on dynamics and relative frequency improved identification of positive clones and elimination of negative clones. and increase the credibility of putative positive clones. Therefore the screening strategy could simplify the subsequent experimental screening and may facilitate the generation of antigen-specific antibodies.
authors.affiliation: Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China.
authors.forename: Ju
authors.initials: J
authors.lastname: Liu
authors.affiliation: Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China.
authors.forename: Ruihua
authors.initials: R
authors.lastname: Li
authors.affiliation: Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China.
authors.forename: Kun
authors.initials: K
authors.lastname: Liu
authors.affiliation: Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China.
authors.forename: Liangliang
authors.initials: L
authors.lastname: Li
authors.affiliation: Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China.
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Zai
authors.affiliation: Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China.
authors.forename: Xiangyang
authors.initials: X
authors.lastname: Chi
authors.affiliation: Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Fu
authors.affiliation: Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China. Electronic address: xujunjie@sina.com.
authors.forename: Junjie
authors.initials: J
authors.lastname: Xu
authors.affiliation: Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China. Electronic address: cw0226@foxmail.com.
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
mesh_terms: D000595:Amino Acid Sequence; D022122:Anthrax Vaccines / Q000737:chemistry; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000941:Antigens / Q000737:chemistry; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D005189:False Positive Reactions; D057809:HEK293 Cells; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry; D007135:Immunoglobulin Variable Region / Q000737:chemistry; D007136:Immunoglobulins / Q000737:chemistry; D008297:Male; D008969:Molecular Sequence Data; D014611:Vaccination
chemical_list: D022122:Anthrax Vaccines; D000911:Antibodies, Monoclonal; D000941:Antigens; C014609:IgK; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D007136:Immunoglobulins
keywords: Antibody repertoire; Antigen-specific human monoclonal antibodies; High-throughput sequencing; Screening strategy