Phage antibodies: filamentous phage displaying antibody variable domains

  title={Phage antibodies: filamentous phage displaying antibody variable domains},
  author={John McCafferty and Andrew D. Griffiths and Gregory Paul Winter and D. J. Chiswell},
NEW ways of making antibodies have recently been demonstrated using gene technology. Immunoglobulm variable (V) genes are amplified from hybridomas or B cells using the polymerase chain reaction, and cloned into expression vectors. Soluble antibody fragments secreted from bacteria are then screened for binding activities (see ref. 1 for review). Screening of V genes would, however, be revolutionized if they could be expressed on the surface of bacteriophage. Phage carrying V genes that encode… 

Making antibodies by phage display technology.

Human antibody fragments with many different binding specificities have been isolated from the same phage repertoire, including haptens, carbohydrates, secreted and cell surface proteins, viral coat proteins, and intracellular antigens from the lumen of the endoplasmic reticulum and the nucleus.

Filamentous Phage Display

This system provides an alternative and powerful tool in generating specific antibodies by using filamentous phage that express combinations of randomly assembled pairs of heavy and light chain genes either as a single chain antibody fragment or fragment antigen binding format.

Clonal Selection and Amplification of Phage Displayed Antibodies by Linking Antigen Recognition and Phage Replication

It is described how the clonal selection mechanisms of the humoral immune resonse can also be mimicked in the phage display system by linking antigen–recognition and phage replication.

Selection and rapid purification of murine antibody fragments that bind a transition-state analog by phage display

A new phage-display vector encoding a polyhistidine tag that has been used for rapid purification of soluble antibody fragments and can provide a large diverse repertoire of candidate catalytic antibodies based on TSA selection and screening.

Isolation of monoclonal antibody fragments from phage display libraries.

The application of this technology to the construction of a phage-displayed single-domain antibody (sdAb) library based on the heavy chain antibody repertoire of a llama, the panning of the library against a peptide antigen and the expression, purification, and characterization of sdAbs isolated by panning are described.

Making antibody fragments using phage display libraries

Using a random combinatorial library of the rearranged heavy and kappa light chains from mice immune to the hapten 2-phenyloxazol-5-one (phOx), diverse libraries of antibody fragments are displayed on the surface of fd phage and elicited many more pairings with strong binding activities.

Phage display technology for human monoclonal antibodies.

This chapter describes the basic protocols for antibody library construction, handling, and selection.

Phage surface presentation and secretion of antibody fragments using an adaptable phagemid vector.

An existing phagemid vector is modified to improve the stability of synthesized soluble antibody fragments and coexpression of chaperonins should be used with caution for library construction due to the expected selection pressure for protease resistant gene III fusions.



Binding activities of a repertoire of single immunoglobulin variable domains secreted from Escherichia coli

Isolated variable domains may offer an alternative to monoclonal antibodies and serve as the key to building high-affinity human antibodies and the name 'single domain antibodies (dAbs)' is suggested for these antigen binding demands.

Filamentous fusion phage: novel expression vectors that display cloned antigens on the virion surface.

Foreign DNA fragments can be inserted into filamentous phage gene III to create a fusion protein with the foreign sequence in the middle. The fusion protein is incorporated into the virion, which

Searching for peptide ligands with an epitope library.

Tens of millions of short peptides can be easily surveyed for tight binding to an antibody, receptor or other binding protein using an "epitope library." The library is a vast mixture of filamentous

Cloning immunoglobulin variable domains for expression by the polymerase chain reaction.

A set of oligonucleotide primers designed to amplify the cDNA of mouse immunoglobulin heavy and light chain variable domains by the polymerase chain reaction are applied to clone and sequence the variable domains of five hybridoma antibodies and to express a mouse-human chimeric antibody that binds to the human mammary carcinoma line MCF-7.

Generation of a large combinatorial library of the immunoglobulin repertoire in phage lambda.

A novel bacteriophage lambda vector system was used to express in Escherichia coli a combinatorial library of Fab fragments of the mouse antibody repertoire. The system allows rapid and easy

Random peptide libraries: a source of specific protein binding molecules.

This type of library makes it possible to identify peptides that bind to proteins (or other macromolecules) that have no previously known affinity for peptides.

Human monoclonals from antigen-specific selection of B lymphocytes and transformation by EBV.

By this method, monoclonal antibodies to both foreign antigens and autoantigens can be prepared from the normal human B-cell repertoire.

Protein engineering of antibody binding sites: recovery of specific activity in an anti-digoxin single-chain Fv analogue produced in Escherichia coli.

  • J. HustonD. Levinson R. Crea
  • Biology, Chemistry
    Proceedings of the National Academy of Sciences of the United States of America
  • 1988
A biosynthetic antibody binding site, which incorporated the variable domains of anti-digoxin monoclonal antibody 26-10 in a single polypeptide chain (Mr = 26,354), was produced in Escherichia coli