抗体抗原实验

The development of antibody-directed enzyme prodrug therapy is reviewed. Antibody-directed enzyme prodrug therapy (ADEPT) generates a cytotoxic drug selectively within deposits of cancer and is designed to give therapy with high levels of tumour selectivity coupled with excep ...

Interaction of antibodies with the neonatal FcRn receptor controls largely the length of their life cycle. Altering the serum persistence of antibodies through modulation of their interaction with the FcRn may be advantageous for achieving high contrast images shortly after appli ...

Antibodies are essential tools for research, diagnostics and therapy. However, sometimes antibodies with a most favourable specificity profile lack sufficient affinity for a desired application. Here, a method is described to increase the affinity of recombinant scFv antibody f ...

In the human immune system, antibodies with high affinities for antigen are created in two stages. A diverse primary repertoire of antibody structures is produced by the combinatorial rearrangement of germline V gene segments, and antibodies are selected from this repertoire by binding ...

This protocol shows how to reduce immunogenicity by removing CD4+ T cell epitopes. These epitopes may limit the efficacy of protein therapeutics by inducing the development of a long-lived humoral immune response. The approach presented here mainly consists in (i) locate the T cell epitopes ...

Guided selection is a method for humanization of pre-existing non-human (for example, mouse) antibodies, based on chain shuffling of V-genes by using phage display technology. Mouse VH and VL domains are used to guide the selection of a human antibody partner and are replaced sequentially or in p ...

Monoclonal antibodies can be humanized by changing murine framework surface residues into those that are typically observed in closely related human counterparts. This technique of resurfacing an antibody is based on the observation that the antigenicity of a protein is determined ...

CDR grafting, or antibody reshaping, is the most clinically validated route to a successful therapeutic humanised monoclonal antibody, and is the process described in this chapter. Accurate determination of the rodent antibody variable region DNA sequences, amplified by RT-PCR, and ...

Antibodies are highly specific, naturally evolved molecules that recognize and eliminate pathogens and tumour associated antigens. The organisation into distinct structural and functional domains facilitates their engineering. Driven by a variety of antibody engineer ...

The history of isotype selection for therapeutic antibodies clearly demonstrates that solid understanding of the biologic effects mediated by immunoglobulin (Ig) subclasses IgG1, IgG2, IgG3, and IgG4 and their variants is a prerequisite for optimal antibody development. The cho ...

Selection of phage-displayed antibodies on antigen-coated immunotubes is a fast and reliable method for the isolation of specific binders from antibody libraries. This protocol describes the selection of antibodies from phagemid libraries and also provides protocols for the ge ...

Phage display of combinatorial antibody libraries is a very efficient method for selecting recombinant antibodies against a wide range of molecules. It has been applied very successfully for the generation of therapeutic antibodies for more than a decade. To increase robustness and re ...

While the selection of antibody fragments from large V-gene libraries via phage display and panning permits the efficient enrichment of pools with antigen-binding activity, the successful identification of individual clones with desired binding specificities constitutes ...

Introduction of exogenous genes into the mouse germline, even followed by tissue specific expression, has been achieved for many transgenes. In addition, gene targeting in embryonic stem cells has silenced undesirable loci. Although several companies offer the production of trans ...

In yeast surface display, yeast cells are exploited to express a protein of interest on their surface, thereby linking it to the encoding DNA within the cell. This display system has become a widely used platform for protein engineering in the past decade, as it confers eukaryotic expression impor ...

Hyperphage is a substitute for M13KO7 helper phage used in antibody phage display. It allows to improve the antibody display efficiency on phage by 2-3 orders of magnitude. This is achieved by forcing the packaging E. coli cell to exclusively use the pIII portion of the antibody fusion protein for its v ...

Phage antibody technology is a powerful approach for generating human antibodies to target antigens. For many therapeutic applications, it is useful to generate antibodies that bind to cell surface receptors in a manner where binding results in internalization of the antibody. This al ...

Phage display is a powerful tool to select antibodies for conformation-specific epitopes from antibody libraries. Based on the M13 pIII phage display technology we describe a cell suspension-based strategy, which allows panning against complex, multimeric, fully functional cell ...

In vitro selections performed by phage display are often performed against a recombinant antigen adsorbed on a solid surface. Passive adsorption of the antigen involves its partial denaturation, which can lead to binders able to strongly recognize the adsorbed antigen but displaying a p ...

A major ‘post-genome’ objective is to fully characterise the human proteome using a comprehensive collection of specific antibodies directed against all human proteins and their variants. To achieve this, substantial reductions in cost and effort per antibody have to be achieved. A con ...