Comparison of three different ELISAs for the detection of recombinant, native and plasma IL-17A.
41802 antibody
500-P07G antibody
Ab, antibody
CI, Confidence Interval
ELISA
ELISA, Enzyme-Linked-Immunosorbent-Assay
E_41802, ELISA using 41802 capture mAb
E_eBio64CAP17, ELISA using eBio64CAP17 capture neutralizing mAb
IL-17A, Interleukin-17A
Inflammation
Interleukin-17A
LCH, Langerhans Cell Histiocytosis
Langerhans Cell Histiocytosis
OD, optical density, E_500-P07G, ELISA using 500-P07G capture polyclonal Ab
PBLs, Peripheral Blood Lymphocytes
PBMCs, peripheral blood mononuclear cells
Plasma
eBio64CAP17 antibody
mAb, monoclonal Antibody
rhIL-17A, recombinant human IL-17A
Journal
MethodsX
ISSN: 2215-0161
Titre abrégé: MethodsX
Pays: Netherlands
ID NLM: 101639829
Informations de publication
Date de publication:
2020
2020
Historique:
received:
09
10
2019
accepted:
13
07
2020
entrez:
11
8
2020
pubmed:
11
8
2020
medline:
11
8
2020
Statut:
epublish
Résumé
Plasma IL-17A detection in Langerhans Cell Histiocytosis (LCH) is currently a source of debate. Indeed, 500-P07G (PeproTech) and 41802 (R&D Systems) anti-IL-17A antibodies have been suspected to recognize nonspecific proteins. To resolve this discrepancy, we set up two new ELISAs by using 41802 or neutralizing eBio64CAP17 (eBioscience) capture monoclonal antibodies that we compared to the commercial PeproTech ELISA kit. The three ELISAs, called E_500-P07G, E_41802 and E_eBio64CAP17, differ in their anti-IL-17A capture antibodies: either polyclonal, monoclonal or neutralizing monoclonal antibodies, respectively. Here, we show that these ELISAs had a similar capacity to specifically detect recombinant or native human IL-17A. However, a significantly lower plasma IL-17A detection was obtained with E_41802 compared to the two other ELISAs. Both E_500-P07G and E_eBio64CAP17 showed similar results. Consequently, we propose that the use of E_500-P07G and E_eBio64CAP17 may ensure more accurate and reliable results in the context of LCH studies. The highest plasma IL-17A levels in LCH patients compared to controls detected by both E_500-P07G and E_eBio64CAP17 ELISAs led us to propose these latter as reference techniques to investigate IL-17A as a potential new biomarker in LCH.•The customization of a new E_eBio64CAP17 ELISA is suitable to detect human IL-17A.•E_eBio64CAP17 ELISA protocol differs only in the anti-IL-17A capture antibody compared to the commercial E_500-P07G PeproTech kit.•Data generated using the E_eBio64CAP17 ELISA are consistent with the PeproTech kit.
Identifiants
pubmed: 32775222
doi: 10.1016/j.mex.2020.100997
pii: S2215-0161(20)30217-X
pii: 100997
pmc: PMC7394768
doi:
Types de publication
Journal Article
Langues
eng
Pagination
100997Informations de copyright
© 2020 The Authors.
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