A novel platform to produce human monoclonal antibodies

Abstract

A new technology has been developed that allows human antibodies to be quickly generated against virtually any antigen. Using a novel process, naïve human B cells are isolated from tonsil tissue and transformed with efficiency up to 85%, thus utilizing a large portion of the human VDJ/VJ repertoire. Through ex vivo stimulation, the B cells class switch and may undergo somatic hypermutation, thus producing a human "library" of different IgG antibodies that can then be screened against any antigen. Since diversity is generated ex vivo, sampling immunized or previously exposed individuals is not necessary. Cells producing the antibody of interest can be isolated through limiting dilution cloning and the human antibody from the cells can be tested for biological activity. No humanization is necessary because the antibodies are produced from human B cells. By eliminating immunization and humanization steps, and screening a broadly diverse library, this platform should reduce both the cost and time involved in producing therapeutic monoclonal antibody candidates.

Investigation of interactions between binding residues and solubility of grafted humanized anti-VEGF IgG antibodies expressed as full-length format in the cytoplasm of a novel engineered E. coli SHuffle strain

“…'Humanized' antibodies typically contain 85-90% human sequence which further reduces the immunogenicity, but the process of humanizing is more technically demanding than the mouse-human fusions used to generate chimeric antibodies. 32 The CDRs on VH and VL of muMAb A.4.6.1 were identied based on sequence hypervariability, in accordance with Kabat and Chothia's denition. 33,34 The CDRs in the light chain were dened as the residues CDR_L1 (24-34), CDR_L2 (50-56), and CDR_L3 (89-97), while the CDRs in the heavy chain were composed of the residues CDR_H1 (26-35), CDR_H2 (50-66), and CDR_H3 (99-112).…”

“…Nonetheless, this type of antibody still carries immunological risk since only the constant regions, representing approximately 70% of the antibody, are human sequences. 32 Second, humanization of muMAb A.4.6.1 was performed by grafting murine CDR sequences onto the Herceptin framework. ‘Humanized’ antibodies typically contain 85–90% human sequence which further reduces the immunogenicity, but the process of humanizing is more technically demanding than the mouse-human fusions used to generate chimeric antibodies.…”

Antibody-Based Immunotherapies as a Tool for Tackling Multidrug-Resistant Bacterial Infections

“…This process leads to antibody activity losses. Conservation of a few rodent sequences in the framework is required to restore the binding [46,47]. The production of fully human monoclonal antibodies was achieved with the development of phage display platforms, and also with the advancements in transgenic mouse platforms [46].…”

Immunization and Immunotherapy Approaches against Pseudomonas aeruginosa and Burkholderia cepacia Complex Infections

“…Nevertheless, they still differ in glycosylation patterns from human antibodies, affecting their half-life and long-term tolerability. To overcome this problem, recently, a technique to utilize human B cells for the production of therapeutic MAbs was developed [ 62 ].…”