Spot Synthesis: An Optimized Microarray to Detect IgE Epitopes

De-Simone, Salvatore G.; Napoleão-Pêgo, Paloma; De-Simone, Thatiane S.

doi:10.1007/978-1-4939-3037-1_20

Cited by 10 publications

(8 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Keeping this consideration in mind, many groups have published extremely exciting results that clearly show that both diagnostic and prognostic indications can be derived from the analysis of the IgE profile directed to peptides of different allergen sources. [77][78][79][80][81][82][83] Of course, the need for large epidemiological studies will be necessary to define rules that can be applied to different populations of patients. However, the possibility of dissecting the immune response mediated by IgE will be included in the fourth diagnostic level of allergic diseases, such as the basophil activation test, in the near future.…”

Section: Available Diagnostic Toolsmentioning

confidence: 99%

A WAO — ARIA — GA2LEN consensus document on molecular-based allergy diagnosis (PAMD@): Update 2020

Ansotegui¹,

Melioli²,

Canonica³

et al. 2020

World Allergy Organization Journal

View full text Add to dashboard Cite

Precision allergy molecular diagnostic applications (PAMD@) is increasingly entering routine care. Currently, more than 130 allergenic molecules from more than 50 allergy sources are commercially available for in vitro specific immunoglobulin E (sIgE) testing. Since the last publication of this consensus document, a great deal of new information has become available regarding this topic, with over 100 publications in the last year alone. It thus seems quite reasonable to publish an update. It is imperative that clinicians and immunologists specifically trained in allergology keep abreast of the new and rapidly evolving evidence available for PAMD@. PAMD@ may initially appear complex to interpret; however, with increasing experience, the information gained provides relevant information for the allergist. This is especially true for food allergy, Hymenoptera allergy, and for the selection of allergen immunotherapy. Nevertheless, all sIgE tests, including PAMD@, should be evaluated within the framework of a patient's clinical history, because allergen sensitization does not necessarily imply clinical relevant allergies.

show abstract

Section: Available Diagnostic Toolsmentioning

confidence: 99%

A WAO — ARIA — GA2LEN consensus document on molecular-based allergy diagnosis (PAMD@): Update 2020

Ansotegui¹,

Melioli²,

Canonica³

et al. 2020

World Allergy Organization Journal

View full text Add to dashboard Cite

show abstract

“…Because we predict from the native Ag, we showed that only a limited number of peptides (compared to the diversity generated in phage-display method) is necessary to map discontinuous epitopes. After synthesis, the functionality of the peptides exploring all the surface of the protein could be assessed in a convenient high-throughput recognition assay, such as the SPOT method [35, 44, 45] or other technologies [46]. If the correct sequence is present in the bank, the Ab should recognize it and this identifies the epitope region on the protein.…”

Section: Discussionmentioning

confidence: 99%

PEPOP 2.0: new approaches to mimic non-continuous epitopes

et al. 2019

View full text Add to dashboard Cite

Background Bioinformatics methods are helpful to identify new molecules for diagnostic or therapeutic applications. For example, the use of peptides capable of mimicking binding sites has several benefits in replacing a protein which is difficult to produce, or toxic. Using peptides is less expensive. Peptides are easier to manipulate, and can be used as drugs. Continuous epitopes predicted by bioinformatics tools are commonly used and these sequential epitopes are used as is in further experiments. Numerous discontinuous epitope predictors have been developed but only two bioinformatics tools have been proposed so far to predict peptide sequences: Superficial and PEPOP 2.0. PEPOP 2.0 can generate series of peptide sequences that can replace continuous or discontinuous epitopes in their interaction with their cognate antibody. Results We have developed an improved version of PEPOP (PEPOP 2.0) dedicated to answer to experimentalists’ need for a tool able to handle proteins and to turn them into peptides. The PEPOP 2.0 web site has been reorganized by peptide prediction category and is therefore better formulated to experimental designs. Since the first version of PEPOP, 32 new methods of peptide design were developed. In total, PEPOP 2.0 proposes 35 methods in which 34 deal specifically with discontinuous epitopes, the most represented epitope type in nature. Conclusion Through the presentation of its user-friendly, well-structured new web site conceived in close proximity to experimentalists, we report original methods that show how PEPOP 2.0 can assist biologists in dealing with discontinuous epitopes. Electronic supplementary material The online version of this article (10.1186/s12859-019-2867-5) contains supplementary material, which is available to authorized users.

show abstract

“…The entire sequence (728aa) of the non-characterized protein nicastrin, deposited in the UNIPROT (Q4DEM3) and a possible component of GSC in T. cruzi , was represented by a continuous series of 144, 14-residue peptides that overlapped preceding and following peptides by 9 residues. The peptides were synthesized in place on a cellulose membrane according to a SPOT synthesis protocol [ 18 ] using an Auto-Spot Robot ASP-222 (Intavis Bioanalytical Instruments AG, Köln, Germany). Coupling reactions were followed by acetylation with acetic anhydride (4%, v / v ) in N , N -dimethylformamide to render peptides unreactive during the subsequent steps.…”

Section: Methodsmentioning

confidence: 99%

Nicastrin-Like, a Novel Transmembrane Protein from Trypanosoma cruzi Associated to the Flagellar Pocket

et al. 2021

Self Cite

View full text Add to dashboard Cite

Nicastrin (NICT) is a transmembrane protein physically associated with the polytypical aspartyl protease presenilin that plays a vital role in the correct localization and stabilization of presenilin to the membrane-bound γ-secretase complex. This complex is involved in the regulation of a wide range of cellular events, including cell signaling and the regulation of endocytosed membrane proteins for their trafficking and protein processing. Methods: In Trypanosoma cruzi, the causal agent of the Chagas disease, a NICT-like protein (Tc/NICT) was identified with a short C-terminus orthologous to the human protein, a large ectodomain (ECD) with numerous glycosylation sites and a single-core transmembrane domain containing a putative TM-domain (457GSVGA461) important for the γ-secretase complex activity. Results: Using the Spot-synthesis strategy with Chagasic patient sera, five extracellular epitopes were identified and synthetic forms were used to generate rabbit anti-Tc/NICT polyclonal serum that recognized a ~72-kDa molecule in immunoblots of T. cruzi epimastigote extracts. Confocal microscopy suggests that Tc/NICT is localized in the flagellar pocket, which is consistent with data from our previous studies with a T. cruzi presenilin-like protein. Phylogenetically, Tc/NICT was localized within a subgroup with the T. rangeli protein that is clearly detached from the other Trypanosomatidae, such as T. brucei. These results, together with a comparative analysis of the selected peptide sequence regions between the T. cruzi and mammalian proteins, suggest a divergence from the human NICT that might be relevant to Chagas disease pathology. As a whole, our data show that a NICT-like protein is expressed in the infective and replicative stages of T. cruzi and may be considered further evidence for a γ-secretase complex in trypanosomatids.

show abstract

Spot Synthesis: An Optimized Microarray to Detect IgE Epitopes

Cited by 10 publications

References 38 publications

A WAO — ARIA — GA2LEN consensus document on molecular-based allergy diagnosis (PAMD@): Update 2020

A WAO — ARIA — GA2LEN consensus document on molecular-based allergy diagnosis (PAMD@): Update 2020

PEPOP 2.0: new approaches to mimic non-continuous epitopes

Nicastrin-Like, a Novel Transmembrane Protein from Trypanosoma cruzi Associated to the Flagellar Pocket

Contact Info

Product

Resources

About