Library E: Highly Optimized Recombinant Antibody Phage-Display Library



    • Using recombinant antibody libraries for drug development has numerous advantages over animal immunization including: faster production times, favorable expression, and not being limited by antigen immunogenicity. However, this approach is limited by the significant effort and resources required to generate such a library.
    • The inventors developed a novel 10^10 member recombinant phage displayed antibody library (library E) based on the highly stable 4D5 scaffold. The library was designed to have biased diversity in the complementary determining regions (CDR) H3 and L3 based off aggregate sequencing and structural data of hits in previous screens with a similar library (library D). All other CDRs consist mainly of tyrosine and serine residues.
    • The library can generate an initial antibody hit against a target of interest, and that hit can then be further mutagenized to produce a lead candidate. The library is rationally designed to increase likelihood of generating an initial hit, and the diversification scheme is highly amenable to further mutagenesis.
    • In proof of concept experiments, library E was screened against the pMHC-bound (HLA-A2) breast cancer antigen HER2/neu-a traditionally challenging target. The lead antibody isolated from the library was highly specific, bound in vitro with an affinity of 59nM, and also bound in vivo breast cancer mouse models.


    Schematic overview of library diversification scheme. Antibody complimentary determining regions (CDRs) are highlighted in color, with the Cα atoms of the varied amino acids shown as spheres. (B) CDR library diversification scheme from the 4D5 scaffold. The CDRs outside of H3 and L3 were primarily changed to Tyrosine (T) or Serine (S). “X” denotes biased amino acid diversity based on aggregate sequences from prior library screens. Parenthesis denotes variable CDR loop length. Sequence of the lead binding clone (E75) is also shown.



    • Rationally designed to maximize efficiency of generating hits
    • Antibodies highly amenable to affinity maturation


    • Therapeutic development
    • Diagnostic development
    • Research reagent development


    • (library transfer)



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