A combinatorial mutagenesis approach for functional epitope mapping on phage-displayed target antigen

Infante, Yanelys Cabrera; Pupo, Amaury; Rojas, Gertrudis

doi:10.4161/mabs.28395

Cited by 18 publications

(18 citation statements)

References 34 publications

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“…While full-length interleukin-2 (IL-2), epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) have been displayed on filamentous phages for mapping purposes, [5][6][7]9 restricting the scanning to the individual domain containing the epitope(s) under investigation is the alternative of choice for larger multi-domain Ags, as it has been done with phage-displayed EGF receptor (EGFR) domain III. 8 In any case, the original folding/antigenicity of the molecule must be preserved.…”

Section: The Importance Of Defining Functional Epitopes On a Targetmentioning

confidence: 99%

“…9 Simultaneous mutagenesis of residues within discrete antigenic regions allowed evaluation of the contribution of each amino acid cluster as a whole to antigenicity. This procedure provides both loss-of-function and gain-of-function data.…”

Section: Deciphering Epitopes With a Diffuse Energetic Landscape Thromentioning

confidence: 99%

“…42 Library design itself (targeting aa that are as far apart as possible and no more than one already known critical residue in each library) was aimed at minimizing the combined effects of several replacements. 44 The recently described combinatorial epitope mapping approach 9 relies on modifying clusters of residues that are in close proximity to each other and have a high likelihood of establishing additive or cooperative interactions with the paratope. Recording the whole picture of these combined effects is thus the goal of this new approach.…”

Section: Deciphering Epitopes With a Diffuse Energetic Landscape Thromentioning

confidence: 99%

“…Recording the whole picture of these combined effects is thus the goal of this new approach. 9 Figure 3. Clusters of residues identified by combinatorial mutagenesis as the epitopes recognized by 2 monoclonal antibodies against EGF.…”

Section: Deciphering Epitopes With a Diffuse Energetic Landscape Thromentioning

confidence: 99%

“…The current article summarizes mapping experiences accumulated during the last 2 y on the bases of phage display of native Ags and a comprehensive mutagenesis exploration of their surfaces to locate the target epitopes. [5][6][7][8][9] The usefulness of such an approach in the context of a plethora of available epitope discovery techniques is discussed. Despite the wide use of phage display to study protein interactions, including epitope mapping, 10 the particular strategy we review here had not been fully exploited until recently.…”

Section: Introductionmentioning

confidence: 99%

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High throughput functional epitope mapping: Revisiting phage display platform to scan target antigen surface

Rojas¹,

Tundidor²,

Infante³

2014

mAbs

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Antibody engineering must be accompanied by mapping strategies focused on identifying the epitope recognized by each antibody to define its unique functional identity. High throughput fine specificity determination remains technically challenging. We review recent experiences aimed at revisiting the oldest and most extended display technology to develop a robust epitope mapping platform, based on the ability to manipulate target-derived molecules (ranging from the whole native antigen to antigen domains and smaller fragments) on filamentous phages. Single, multiple and combinatorial mutagenesis allowed comprehensive scanning of phage-displayed antigen surface that resulted in the identification of clusters of residues contributing to epitope formation. Functional pictures of the epitope(s) were thus delineated in the natural context. Successful mapping of antibodies against interleukin-2, epidermal growth factor and its receptor, and vascular endothelial growth factor showed the versatility of these procedures, which combine the accuracy of site-directed mutagenesis with the high throughput potential of phage display.

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Section: The Importance Of Defining Functional Epitopes On a Targetmentioning

confidence: 99%

Section: Deciphering Epitopes With a Diffuse Energetic Landscape Thromentioning

confidence: 99%

Section: Deciphering Epitopes With a Diffuse Energetic Landscape Thromentioning

confidence: 99%

Section: Deciphering Epitopes With a Diffuse Energetic Landscape Thromentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

High throughput functional epitope mapping: Revisiting phage display platform to scan target antigen surface

Rojas¹,

Tundidor²,

Infante³

2014

mAbs

Self Cite

View full text Add to dashboard Cite

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Epitope Mapping: B ‐Cell Epitopes

Pulzová

2018

Encyclopedia of Life Sciences

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Antigen–antibody interaction is a key event in humoral immune response to invading pathogen. All of the macromolecular components of living organism (proteins, carbohydrates, lipids and nucleic acids) are potential antigens. A specific antibody recognises antigen at discrete regions known as antigenic determinants or B‐cell epitopes. Most of the antigen surface may become part of epitopes after recognition with specific antibodies; however, the exact selection mechanism of why certain regions become B‐cell epitopes is not fully understood. The epitope mapping is the process of identification and characterisation of the antibody‐binding site on the antigen, although the term is also applied more broadly to receptor–ligand interactions unrelated to the immune system. With an ever‐increasing number of available pathogen genomes, mapping of epitopes becomes the central issue of vaccine development. Precise identification of epitope is crucial for understanding the molecular basis of immunity and autoimmunity and allows to replace an antigen in the immunisation, antibody production and serodiagnosis. Key Concepts Virtually any surface‐exposed residue of antigen may become part of epitope after interactions with the antibody. The exact selection mechanism governing the antibody recognition of the corresponding epitope is not known. The experimental methods developed to identify the epitopes depend on the unique features of the reaction between an antigen and its complementary antibody. Structural methods interpret the protein structure comprising residues in direct contact with an antibody. Functional methods identify and characterise residues important for binding within structurally defined epitope and are based on detection of binding of antibody to antigen fragments. Identification of B‐cell epitopes is extensively employed in the development of diagnostic tests, therapeutics and vaccines. The epitope‐based subunit vaccines became a promising alternative to traditional vaccines; however, low immunogenicity and low affinity of raised antibodies present major drawbacks of these types of vaccines.

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Application of phage peptide display technology for the study of food allergen epitopes

Chen

Dreskin

2017

Molecular Nutrition Food Res

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Phage peptide display technology has been used to identify IgE-binding mimotopes (mimics of natural epitopes) that mimic conformational epitopes. This approach is effective in the characterization of those epitopes that are important for eliciting IgE-mediated allergic responses by food allergens and those that are responsible for cross-reactivity among allergenic food proteins. Application of this technology will increase our understanding of the mechanisms whereby food allergens elicit allergic reactions, will facilitate the discovery of diagnostic reagents and may lead to mimotope-based immunotherapy.

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A combinatorial mutagenesis approach for functional epitope mapping on phage-displayed target antigen

Cited by 18 publications

References 34 publications

High throughput functional epitope mapping: Revisiting phage display platform to scan target antigen surface

High throughput functional epitope mapping: Revisiting phage display platform to scan target antigen surface

Epitope Mapping: B ‐Cell Epitopes

Application of phage peptide display technology for the study of food allergen epitopes

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