Diverse human VH antibody fragments with bio-therapeutic properties from the Crescendo Mouse

• Published in: New biotechnology 2020-03, Vol.55, p.65-76
• Main Authors: Teng, Yumin, Young, Joyce L., Edwards, Bryan, Hayes, Philip, Thompson, Lorraine, Johnston, Colette, Edwards, Carolyn, Sanders, Yun, Writer, Michele, Pinto, Debora, Zhang, Yanjing, Roode, Mila, Chovanec, Peter, Matheson, Louise, Corcoran, Anne E., Fernandez, Almudena, Montoliu, Lluis, Rossi, Beatrice, Tosato, Valentina, Gjuracic, Kresimir, Nikitin, Dmitri, Bruschi, Carlo, McGuinness, Brian, Sandal, Thomas, Romanos, Mike
• Format: Article
• Language: English
• Subjects: Antibodies; B-lymphocytes; bacteriophages; Chains; Crescendo Mouse; Display devices; E coli; Escherichia coli; Fragments; genes; genetically modified organisms; heavy chain antibody; Humabody®+VH; humans; Immunization; immunoglobulins; Interleukin 1; Loci; Lymphocytes B; mice; Phage display; Phages; Somatic hypermutation; Variable domain; VH domain
• ISSN: 1871-6784; 1876-4347
• DOI: 10.1016/j.nbt.2019.10.003

[Display omitted] •Crescendo Mouse produces heavy chain only antibodies with fully human variable domains.•Triple knock out background is essential for in vivo maturation of HC-only antibody.•Crescendo Mouse shows robust responses to immunisation generating high affinity Humabody® VH against therapeutic targets.•Humabody® VH fragments possess desirable biophysical properties for therapeutic development. We describe the ‘Crescendo Mouse’, a human VH transgenic platform combining an engineered heavy chain locus with diverse human heavy chain V, D and J genes, a modified mouse Cγ1 gene and complete 3’ regulatory region, in a triple knock-out (TKO) mouse background devoid of endogenous immunoglobulin expression. The addition of the engineered heavy chain locus to the TKO mouse restored B cell development, giving rise to functional B cells that responded to immunization with a diverse response that comprised entirely ‘heavy chain only’ antibodies. Heavy chain variable (VH) domain libraries were rapidly mined using phage display technology, yielding diverse high-affinity human VH that had undergone somatic hypermutation, lacked aggregation and showed enhanced expression in E. coli. The Crescendo Mouse produces human VH fragments, or Humabody® VH, with excellent bio-therapeutic potential, as exemplified here by the generation of antagonistic Humabody® VH specific for human IL17A and IL17RA.