Comparison of three different ELISAs for the detection of recombinant, native and plasma IL-17A

Mohamad Bachar Ismail; Selma Olsson Åkefeldt; Magda Lourda; Désirée Gavhed; Rémi Gayet; Maurizio Aricò; Jan-Inge Henter; Christine Delprat; Hélène Valentin

doi:10.1016/j.mex.2020.100997

Comparison of three different ELISAs for the detection of recombinant, native and plasma IL-17A

MethodsX. 2020 Jul 16:7:100997. doi: 10.1016/j.mex.2020.100997. eCollection 2020.

Authors

Mohamad Bachar Ismail¹, Selma Olsson Åkefeldt^{2

3}, Magda Lourda^{2

4}, Désirée Gavhed², Rémi Gayet⁵, Maurizio Aricò^{6

7}, Jan-Inge Henter^{2

3}, Christine Delprat^{8

9}, Hélène Valentin⁹

Affiliations

¹ Laboratoire Microbiologie Santé et Environnement, Doctoral School of Sciences and Technology, Faculty of Public Health, Lebanese University, Tripoli, Lebanon.
² Childhood Cancer Research Unit, Department of Women's and Children's Health, Karolinska Institutet, Karolinska University Hospital, Stockholm, Sweden.
³ Theme of Children's and Women's Health, Karolinska University Hospital, Stockholm, Sweden.
⁴ Center for Infectious Medicine, Department of Medicine, Karolinska Institutet, Stockholm, Sweden.
⁵ Faculté de Médecine, Groupe sur l'Immunité des Muqueuses et Agents Pathogènes (GIMAP), 10 rue de la Marandière, 42270 Saint Priest en Jarez, France.
⁶ Azienda Ospedaliero-Universitaria A. Meyer Children Hospital, Firenze, Italy.
⁷ Ospedale Pediatrico Giovanni XXIII Azienda Ospedaliero Universitaria Consorziale Policlinico Bari.
⁸ UnivLyon, Université Claude Bernard Lyon 1, Villeurbanne, France.
⁹ INSERM U1052, CNRS UMR5686, Cancer Research Center of Lyon (CRCL), Lyon, France.

Abstract

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.

Keywords: 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.