Delineating the functional map of the interaction between nimotuzumab and the epidermal growth factor receptor

Tundidor, Yaima; García-Hernández, Claudia Patricia; Pupo, Amaury; Infante, Yanelys Cabrera; Rojas, Gertrudis

doi:10.4161/mabs.28915

Cited by 17 publications

(50 citation statements)

References 56 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…While lowlevel random diversification of phagemid-cloned Ag gene by error-prone PCR was initially performed, 25,26 resembling the usual yeast display-based mapping procedures, detailed information about the residues contributing to epitope formation has been recently obtained through comprehensive mutagenesis scanning of the role of solvent-exposed side chains within a discrete surface patch. [5][6][7][8] Once the target Ag is phage-displayed and a putative relevant antigenic region within it is identified through any available method, focused exploration of such region can result in a very high local sequence space coverage (Fig. 1).…”

Section: The Importance Of Defining Functional Epitopes On a Targetmentioning

confidence: 99%

“…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. All the mentioned Ags have disulfide bonds (from one in IL-2 to 5 in EGFR domain III).…”

Section: The Importance Of Defining Functional Epitopes On a Targetmentioning

confidence: 99%

“…Commonly used phagemids are easily produced as the required single-strand DNA mutagenesis templates. Other site-directed mutagenesis procedures have been used to obtain Ag-derived variants, 6 but the extreme simplicity and speed of Kunkel mutagenesis 8 Functional epitopes have been identified on phage-displayed interleukin-2 (IL-2), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF) and EGF receptor (EGFR) domain III. The initial antigenic regions to be characterized were defined through different approaches.…”

Section: The Importance Of Defining Functional Epitopes On a Targetmentioning

confidence: 99%

“…In such cases, phage purification/concentration is required and can be performed through a simple precipitation with polyethyleneglycol, 38 still allowing the evaluation of hundreds of mutated variants. 8 A proper judgment of sitedirected mutagenesis results requires careful examination. Since conclusions are supported by loss-ofbinding data, it is crucial to rule out gross folding defects affecting protein display or conformation.…”

Section: The Importance Of Defining Functional Epitopes On a Targetmentioning

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%

See 4 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

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.

show abstract

Section: The Importance Of Defining Functional Epitopes On a Targetmentioning

confidence: 99%

Section: The Importance Of Defining Functional Epitopes On a Targetmentioning

confidence: 99%

Section: The Importance Of Defining Functional Epitopes On a Targetmentioning

confidence: 99%

Section: The Importance Of Defining Functional Epitopes On a Targetmentioning

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

show abstract

Analysis of EGFR binding hotspots for design of new EGFR inhibitory biologics

Tydings,

Singh,

Smith

et al. 2024

Protein Science

View full text Add to dashboard Cite

The epidermal growth factor (EGF) receptor (EGFR) is activated by the binding of one of seven EGF‐like ligands to its ectodomain. Ligand binding results in EGFR dimerization and stabilization of the active receptor conformation subsequently leading to activation of downstream signaling. Aberrant activation of EGFR contributes to cancer progression through EGFR overexpression/amplification, modulation of its positive and negative regulators, and/or activating mutations within EGFR. EGFR targeted therapeutic antibodies prevent dimerization and interaction with endogenous ligands by binding the ectodomain of EGFR. However, these antibodies have had limited success in the clinic, partially due to EGFR ectodomain resistance mutations, and are only applicable to a subset of patients with EGFR‐driven cancers. These limitations suggest that alternative EGFR targeted biologics need to be explored for EGFR‐driven cancer therapy. To this end, we analyze the EGFR interfaces of known inhibitory biologics with determined structures in the context of endogenous ligands, using the Rosetta macromolecular modeling software to highlight the most important interactions on a per‐residue basis. We use this analysis to identify the structural determinants of EGFR targeted biologics. We suggest that commonly observed binding motifs serve as the basis for rational design of new EGFR targeted biologics, such as peptides, antibodies, and nanobodies.

show abstract

FITC-conjugated single chain Nimotuzumab can specifically recognize and enter EGFR-overexpressing cells

Hamidi

Safdari

Arabi

2022

Int J Pept Res Ther

View full text Add to dashboard Cite

Delineating the functional map of the interaction between nimotuzumab and the epidermal growth factor receptor

Cited by 17 publications

References 56 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

Analysis of EGFR binding hotspots for design of new EGFR inhibitory biologics

FITC-conjugated single chain Nimotuzumab can specifically recognize and enter EGFR-overexpressing cells

Contact Info

Product

Resources

About