Immune phage

Vaughan, T. J., Williams, A. J., Pritchard, K., Osbourn, J. K., Pope, A. R., Eamshaw, J. C., McCafferty, J., Hodits, R. A., Wilton, J., and Johnson, K. S. (1996). Human antibodies with sub-nanomolar affinities isolated from a large non-immunized phage display library. Nat. Biotechnol. 14, 309-314. 

To circumvent some of the limitations of direct immunization, phage display technology has been applied to the preparation of fully human MABs. Gene libraries of cDNA from nonimmune or immunized donor lymphocytes are expressed in bacteriophages. The bacteriophages display functional antibody fragments and can... 

Chan SW, Bye JM, Jackson P, Allain JP, Human recombinant antibodies specific for hepatitis C virus core and envelope E2 peptides from an immune phage display library, J. Gen. Virol., 77 2531-2539, 1996. 

The affinity of antibodies selected from naive, synthetic, or even immune phage libraries is typically sufficient for use as a research reagent, but too low for some specific therapeutic applications such as viral neutralization or tumor imaging. For many applications, affinity maturation can be bypassed completely by the construction of multivalent molecules. If this is not sufficient selected antibody clones may be affinity matured (see Figure 18.13). 

Sheets MD, et al., Efficient construction of a large non-immune phage antibody library the production of panels of high affinity human single-chain antibodies to protein antigens, Proc. Natl. Acad. Sci. USA., 95 6157-6162, 1998. 

Vaughan, T.J., et al., Human antibodies with sub-nanomolar affinities isolated from a large non-immunized phage display library, Nat. BiotechnoL, 14, 309, 1996. 

Marks, J. D., Hoogenboom, H. R., Bonnert, T. P., McCafferty, J., Griffiths, D., and Winter, G. (1991). By-passing immunization. Human antibodies from V-gene libraries displayed on phage. J. Mol. Biol. 222, 581-597. 

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. 

More recently, Janda has described the production of a galactopyranosidase antibody in response to hapten [96]. This was designed to accommodate several features of the transition state for glycoside hydrolysis notably a flattened half-chair conformation and substantial sp2 character at the anomeric position. Some 100 clones were isolated in response to immunization with [96] and used to generate a cDNA library for display on the surface of phage (Appendix entry 7.3) (Janda et al., 1997). Rather than proceed to the normal screening for turnover, Janda then created a suicide substrate system to trap the catalytic species. 

Figure 28-11 Genetic and physical map of the X phage genome. After Szybalski. See Honigman et al.255 for a more detailed diagram of the immunity region. The gene for the lambda repressor is labeled C[.

An artificial immune system was constructed based on phage display to make antibodies without using rodents and hybridoma technology.23,26,27 The gene III... 

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