Mining human antibody repertoires

Transpo-mAb display: Transposition-mediated B cell display and functional screening of full-length IgG antibody libraries

“…These include, for instance, phage display, retroviral display, bacterial display, yeast display and various mammalian cell display technologies, in combination with solid surface binding (panning) or other enrichment techniques. 2,3 While phage, 4-6 prokaryotic 7,8 and ribosome/mRNA display 9,10 systems have been established and are widely adopted in the biotechnology industry and in academia for the identification of antibody fragments, they suffer from a variety of limitations, including the inability to express full-length antibodies, the absence of natural post-translational modifications, the lack of proper folding by vertebrate chaperones, and an artificially enforced heavy and light chain combination. Therefore, the "reformatting" of antibody fragments into full-length antibodies followed by manufacturing in mammalian cells frequently results in molecules with unfavorable biophysical properties (e.g., low stability, tendency to aggregate, diminished affinity).…”

Multiplexed screening of natural humoral immunity identifies antibodies at fine specificity for complex and dynamic viral targets

“…9 An appreciation of the complexity and diversity of antibody responses in the human population and the resulting rarity of broadly protective memory B cell clones led to the development PAPer TyPe rePorT of a number of human antibody cloning technologies. 10,11 Herein, we employed a multiplexed screening process to enable an in-depth characterization of the specificity of naturally occurring antibodies secreted from single memory B cells. Deeming multiplexing a critical component to discovering antiviral antibodies with cross-clade activity, we counteracted the associated rapid drop in hit frequency with high throughput and miniaturized assay technologies.…”

High Affinity Maturated Human Antibodies from Naïve and Synthetic Antibody Repertoires

“…The kappa light chain locus is found in chromosome 2 with the V K , J K and C K gene segments. The lambda LC locus with the V λ , J λ , and C λ gene segments are found on chromosome 22 [37].…”