Display naive antibody

Willats WGT, Gilmartin PM, Mikkelsen JD, Knox JP. Cell wall antibodies without immunization generation and use of de-esterified homogalacturonan block-specific antibodies from a naive phage display library. Plant J. 1999 18 57-65. 

To achieve surface display, five of the Ml3 coat proteins have been used in fusion to foreign protein fragments. In the most widespread system the antibody is coupled to the N-terminus or second domain of the minor coat protein pill (11,12,14). The naive function of the three to five copies of the pill, in particular their N-terminal domain, is to provide binding of the phage to the f-pili of E. coli to initiate infection (39). The major coat protein (pVIII) has been used as an alternative fusion partner, with only very few successes reported in the past decade (40). This fusion technique is more useful for the display of short peptides (41,42). Fusions to pVI have been tried, but not yet with antibodies (43). pVII and pIX were... 

In ribosome display, the physical link between genotype and phenotype is accomplished by mRNA-ribosome—protein complexes, which are directly used for selection. If a library of different mRNA molecules is translated, a protein library results in which each protein is produced from its own mRNA and remains connected to it. Since these complexes of the proteins and their encoding mRNAs are stable for several days under the appropriate conditions, very stringent selections can be performed. As all steps of ribosome display are carried out in vitro, reaction conditions of the individual steps can be tailored to the requirements of the protein species investigated, as well as the objectives of the selection or evolution experiment. Application of ribosome display has produced scFv fragments of antibodies with affinities in the picomolar range from libraries prepared from immunized mice (Hanes et al., 1998) and more recently from a naive, completely synthetic library (Hanes et al., 2000), and has been used to evolve improved off-rates and stability (Jermutus et al., 2000). 

This discussion has focused on the desire to create a one-pot naive phage antibody library. However, phage display has also been used very effectively to isolate binding antibodies when libraries are made from immunized experimental animals [32], or from naturally immunized humans [7, 30] in these cases the V-gene source being either spleen or peripheral-blood lymphocytes. 

Hanes, J., SchafFitzel, C., Knappik, A., Pliick-thun, A. Picomolar affinity antibodies from a fully synthetic naive library selected and evolved by ribosome display. Nat. Biotechnol. 2000, 38, 1287-1292. 

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