TY - JOUR
T1 - Complete humanization of the mouse immunoglobulin loci enables efficient therapeutic antibody discovery
AU - Lee, E-Chiang
AU - Liang, Qi
AU - Ali, Hanif
AU - Bayliss, Luke
AU - Beasley, Alastair
AU - Bloomfield-Gerdes, Tara
AU - Bonoli, Laura
AU - Brown, Richard
AU - Campbell, Jamie
AU - Carpenter, Adam
AU - Chalk, Sara
AU - Davis, Alison
AU - England, Nick
AU - Fane-Dremucheva, Alla
AU - Franz, Bettina
AU - Germaschewski, Volker
AU - Holmes, Helen
AU - Holmes, Steve
AU - Kirby, Ian
AU - Kosmac, Miha
AU - Legent, Anais
AU - Lui, Hui
AU - Manin, Anais
AU - O'Leary, Siobhan
AU - Paterson, Jemima
AU - Sciarrillo, Rocco
AU - Speak, Anneliese
AU - Spensberger, Dominik
AU - Tuffery, Laura
AU - Waddell, Nikole
AU - Wang, Wei
AU - Wells, Sophie
AU - Wong, Vivian
AU - Wood, Andrew
AU - Owen, Michael J
AU - Friedrich, Glenn A
AU - Bradley, Allan
PY - 2014/4
Y1 - 2014/4
N2 - 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.
AB - 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.
KW - Amino Acid Sequence
KW - Animals
KW - Antibodies, Monoclonal/genetics
KW - B-Lymphocytes/immunology
KW - Chromosomes, Artificial, Bacterial/genetics
KW - Female
KW - Genetic Engineering/methods
KW - Humans
KW - Immunoglobulin Variable Region/genetics
KW - Male
KW - Mice
KW - Mice, Transgenic
KW - Molecular Sequence Data
KW - Transgenes/genetics
U2 - 10.1038/nbt.2825
DO - 10.1038/nbt.2825
M3 - Article
C2 - 24633243
VL - 32
SP - 356
EP - 363
JO - Nature Biotechnology
JF - Nature Biotechnology
SN - 1087-0156
IS - 4
ER -