Humira

MabThera^® (rituximab reference product) was from F. Hoffmann‐La Roche AG (Basel, Switzerland) and provided as a 10.0 mg/mL aqueous solution containing sodium citrate dihydrate, sodium chloride, and polysorbate 80, at pH 6.5. Reditux^TM (10.0 mg/mL; copy product) was from Dr. Reddy´s Laboratories Ltd. (Hyderabad, India) and provided in the same formulation buffer as MabThera^®. Humira^® (adalimumab) drug product (48.5 mg/mL, pH 5.2) was from AbbVie Inc. (Lake Bluff, Il, USA), containing mannitol, citric acid monohydrate, sodium citrate, sodium dihydrogen phosphate dihydrate, disodium phosphate dihydrate, sodium chloride, polysorbate 80, and sodium hydroxide. All antibodies were stored below −60°C.

PANTS is a UK-wide, multicentre, prospective observational cohort study reporting the treatment failure rates of the anti-TNF drugs infliximab (originator, Remicade [Merck Sharp & Dohme] and biosimilar, CT-P13 [Celltrion]), and adalimumab (Humira [AbbVie]) in 1610 anti-TNF naive CD patients. The study design has been described in detail previously.^{8 (link),22 (link)} In brief, patients were recruited at the time of first anti-TNF exposure between February 2013 and June 2016, and evaluated for 12 months or until drug withdrawal. Eligible patients were aged ≥6 years with evidence of active luminal CD involving the colon and/or small intestine. Four major study visits were scheduled at week 0 [baseline], week 14 [post-induction], week 30, and week 54. Additional visits were scheduled at treatment failure or study exit. At baseline, clinical and demographic data were recorded, including sex, ethnicity, BMI, smoking status, age at diagnosis, disease duration, Montreal classification, prior medical and drug history, and previous CD-related surgeries. At every visit, disease activity score, weight, current therapy, and adverse events were recorded.^{8 (link)}

Concentration of anti-TNF-α mAb was measured by ELISA using IgG-ELISA-BEST kit (Vector-Best, Russia), calibrated for Humira (AbbVie, USA). SDS-PAGE electrophoresis under reducing conditions was performed according to the standard procedure in a 12 % Laemmli gel followed by staining with Coomassie-R250 to check the ratio of heavy and light chains in the supernatant.

For 18 COVID-19 patients, one milliliter of whole blood taken at D0 was pretreated with different molecules for 6 h at 37°C, followed by stimulation with immune ligands on single lyophilized spheres (LyoSphere^TM, Qiagen), as described under Blood collection and cytokine assay. The molecules used were those commonly administered to COVID-19 patients (33 (link)–43 (link)): hydroxychloroquine (100 μM, Inresa), anti-IL6 Tocilizumab (100 μg/mL, RoActemra, Roche), methylprednisolone (20 μg/mL, Mylan), anti-TNFα Adalimumab (10 μg/mL, Humira, AbbVie), recombinant human IL-2 (6 ng/mL, Sigma), recombinant human IFN-alpha (100 ng/mL, Sigma) and Nivolumab (1 μg/mL, Opdivo, Bristol Myers Squibb).