Phage display antibody library

Malmborg AC, Duenas M, Ohlin M, Soderlind E, Borrebaeck C, Selection of binders from phage displayed antibody libraries using the BIAcore biosensor, J. Immunol. Meth., 198 51-57, 1996. 

Sidhu, S. S., B. Li, Y. Chen, F. A. Fellouse, C. Eigenbrot and G. Fuh (2004). Phage-displayed antibody libraries of synthetic heavy chain complementarity determining regions. JMol Biol 338(2) 299-310. 

Phage display antibody libraries are relatively new and have been used in a few P450 applications to date (D.S. Keeney personal communication). These have a number of advantages, including jjotential selectivity due to the large number of potential antibodies in libraries, the ability to avoid animal protocols, the immediate availability of libraries (as opposed to waiting on animals to develop antibodies), the consistency of reproduction of the proteins propagated in bacterial systems, and the ability to include a second epitope tag for recovery, etc. 

The ability to produce/isolate specific antibodies to a number of different proteins from the phage display library offer a potential to provide antibodies for genome scale application of protein chips (Lee and Mrksich, 2002). Phage display of combinatorial antibody library (subsection 13.7.5) is uniquely capable of providing specific monoclonal antibodies in the number and at the rate required for bioinfoimatics research. Table 16.16 lists some commercial sources of phage display antibody libraries. 

MAbs directed against the C. neoformans polysaccharide are closely related, and some differ only in a few amino acid residues [142,143]. This fine specificity is related to their protective efficacy. A highly protective IgGl mAh was used to screen phage-displayed peptide libraries, and several binding peptide sequences were identified [144]. The dodecapeptide GLQYTPSWMLVG bound to the mAh with a Kd of 295 nM, measured by surface plasmon resonance (SPR) and inhibited binding of the mAh to the polysaccharide. The dodecapeptide was able to induce only a weak antipolysaccharide antibody response [145]. 

A major source of human antibodies are phage display libraries, which are constructed from various genetic sources. Antibodies are expressed as scFV and Fab antibody fragments using various vector systems. This review offers a comprehensive overview of M13 phage display antibody vectors and discusses their applications. 

In some cases, functional assays have been used to screen phage-display peptide libraries. For example, a phage-display hexapeptide library was constructed with an epitope tag distal (N-terminus) to screen for peptide substrates of a specific protease. All the phages were captured by an immobilized anti-epitope antibody. After incubation with a tissue plasminogen activator (tPA), phages that expressed a peptide substrate for tPA were released for subsequent rounds of selection. A similar approach was applied to discover peptide substrates for HIV-1 protease. 

There are two general methods for identifying antibody epitopes (1) evaluate peptides whose composition is based on the sequence of the native protein, or (2) evaluate peptides that are selected from a random combinatorial peptide library. The former is only effective if the epitope is composed of a linear sequence of amino acids in the native protein sequence. If it is, then analysis of overlapping peptides from the native protein is a simple method for identifying antibody epitopes. Each peptide is tested for immunoreactivity to the antibody. Those peptides that are immunoreactive contain the epitope. The other method of epitope identihcation, selection from a random combinatorial library, will identify peptides that represent both linear and conformationally dependent epitopes. The drawback of this method is that biopanning from a random combinatorial peptide library is more time consuming. Phage-displayed peptide libraries have previously been used for epitope identification in this context. ... 

SchmaljohnC, Cui Y, Kerby S,et al. (1999). Production and characterization of human monoclonal antibody Fab fragments to vaccinia virus from a phage-display combinatorial library. Virol. 258 189-200. 

Davies E. L., Smith J. S., Birkett C. R., Manser J. M., Anderson-Dear D. V., and Young J. R., Selection of specific phage-display antibodies using libraries derived from chicken immunoglobulin genes, J. Immunol. Meth., 186, 125-135, 1995. 

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