Complete humanization of the mouse immunoglobulin loci enables efficient therapeutic antibody discovery

• Published in: Nature biotechnology Vol. 32; no. 4; pp. 356 - 363
• Main Authors: Lee, E-Chiang, Liang, Qi, Ali, Hanif, Bayliss, Luke, Beasley, Alastair, Bloomfield-Gerdes, Tara, Bonoli, Laura, Brown, Richard, Campbell, Jamie, Carpenter, Adam, Chalk, Sara, Davis, Alison, England, Nick, Fane-Dremucheva, Alla, Franz, Bettina, Germaschewski, Volker, Holmes, Helen, Holmes, Steve, Kirby, Ian, Kosmac, Miha, Legent, Anais, Lui, Hui, Manin, Anais, O'Leary, Siobhan, Paterson, Jemima, Sciarrillo, Rocco, Speak, Anneliese, Spensberger, Dominik, Tuffery, Laura, Waddell, Nikole, Wang, Wei, Wells, Sophie, Wong, Vivian, Wood, Andrew, Owen, Michael J, Friedrich, Glenn A, Bradley, Allan
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.04.2014; Nature Publishing Group
• Subjects: 42/100; 42/41; 631/154/51/1568; 631/1647/767/1972; 631/250/248; 631/61/51/1568; 64/110; 82/1; 82/47; Agriculture; AIDS vaccines; Amino Acid Sequence; Animal genetic engineering; Animals; Antibodies; Antibodies, Monoclonal - genetics; Antibodies, Monoclonal - immunology; Antigenic determinants; Antigens; B cells; B-Lymphocytes - immunology; B-Lymphocytes - physiology; Bioinformatics; Biomedical Engineering/Biotechnology; Biomedicine; Biopharmaceuticals; Biotechnology; Chromosomes, Artificial, Bacterial - genetics; Embryonic stem cells; Female; Genes; Genetic engineering; Genetic Engineering - methods; Genomes; Genomics; Health aspects; House mouse; Humans; Identification and classification; Immune response; Immune system; Immunization; Immunoglobulin Variable Region - genetics; Immunoglobulins; Isolation; Life Sciences; Male; Mice; Mice, Transgenic; Molecular Sequence Data; Monoclonal antibodies; Mutation; Properties; Rodents; Stem cells; Technology; Transgenes - genetics; Vaccines; Viral antibodies; United States; United Kingdom
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/nbt.2825

Mice engineered to express the entire human variable immunoglobulin region enable rapid isolation of high-affinity antibodies specific for antigens of interest. If immunized with an antigen of interest, transgenic mice with large portions of unrearranged human immunoglobulin loci can produce fully human antigen-specific antibodies; several such antibodies are in clinical use. However, technical limitations inherent to conventional transgenic technology and sequence divergence between the human and mouse immunoglobulin constant regions limit the utility of these mice. Here, using repetitive cycles of genome engineering in embryonic stem cells, we have inserted the entire human immunoglobulin variable-gene repertoire (2.7 Mb) into the mouse genome, leaving the mouse constant regions intact. These transgenic mice are viable and fertile, with an immune system resembling that of wild-type mice. Antigen immunization results in production of high-affinity antibodies with long human-like complementarity-determining region 3 (CDR3H), broad epitope coverage and strong signatures of somatic hypermutation. These mice provide a robust system for the discovery of therapeutic human monoclonal antibodies; as a surrogate readout of the human antibody response, they may also aid vaccine design efforts.