Triton x 405

Ovaries of female worms were isolated using the combined detergents/enzyme-based organ isolation protocol [42 (link)]. In short, isolated adult females (about 50 each) were transferred into 2 ml-reaction vessels and washed twice with 2 ml of non-supplemented M199-medium at room temperature. The medium was removed, and 500 μl of tegument solubilisation (TS)-solution was added (0.1% of each following compounds in DEPC (diethylpyrocarbonate)/PBS (phosphate-buffered saline): Brij 35 (Roth), Nonidet P40-Substrate (Fluka), Tween80 (Sigma) and TritonX-405 (Sigma), pH 7.2–7.4) followed by incubation in a thermal shaker (TS-100, Biosan) for 5 min at 1,200 rpm at 37°C. Shaking was repeated twice, and the solution was replaced after each cycle. Then the worms were rinsed three times with M199 and subsequently treated with elastase (300 μl elastase solution: 5 U/ml in M199; Sigma) at 37°C and 650 rpm in the thermal shaker to release the ovaries. Digestion was monitored by bright-field microcopy (Leica) and stopped when the gonads were released from the disrupted and digested worm carcasses. Finally, the gonads were manually collected by pipetting and transferred into supplemented M199 medium.

Metakaolin (MetaMax, BASF; 53.0 weight-% SiO₂ and 44.5 weight-% Al₂O₃) was used as an aluminosilicate precursor. The alkaline solution was prepared by mixing a sodium silicate solution (Merck; molar SiO₂/Na₂O ≈ 3.5, water content ≈ 64 weight-%) with sodium hydroxide pellets (VWR Chemicals) to result in a molar ratio of SiO₂/Na₂O = 1.2.
Hydrogen peroxide (Honeywell; 30%, w/v) was used as a blowing agent in the direct foaming method. Surfactants employed in the study were Triton X-114 (Acros Organics; 100% solution), Triton X-100 (Sigma-Aldrich; 100% solution), Triton X-405 (Sigma-Aldrich; 70% solution), sodium dodecyl sulfate (SDS; Sigma-Aldrich; ≥98.5 weight-% solid), and cetyltrimethylammonium bromide (CTAB; AMRESCO; ≥ 99 weight-% solid; Supporting Information Fig. S4 for additional information). Polyethylene glycol (PEG-1000; Merck) was used to control the rheology in 3D printing.
Metal modification was conducted using Cu(NO₃)₂·3H₂O (VWR Chemicals; ≥97 weight-%) and AgNO₃ (VWR Chemicals; ≥99.5 weight-%); bentonite modified with metallic Ag (Ag⁰) and zinc pyrithione (ArgiBlock 001.ZnPy, Laboratorios Argenol; Ag content 1.93 weight-%, d₅₀ = 8.12 μm); and colloidal Ag solution (Laboratorios Argenol, Ag content 3051 ppm). The colloidal Ag solution consisted of Ag⁰ particles suspended in water.

The 4-aminophenylacetic acid 98%, the N-Hydroxysuccinimide (NHS), and the 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) were purchased from Acros Organics. Sodium nitrite was acquired from Fisher Scientific. Triton X-405 and phosphate buffer saline tablet were acquired from Sigma, ethanolamine from Fluka analytical, and hydrochloric acid 37% (HCl) from VWR, France. Iron (II) chloride tetrahydrate (FeCl₂·4H₂O), iron (III) chloride hexahydrate (FeCl₃·6H₂O), and oleic acid were purchased from Merck. Octane, ammonium hydroxide, and chlorhydric acid were from Prolabo. The preparation of the described bioconjugates and antibodies (5-[4-(amino)phenylsulfonamide]-5-oxopentanoic acid (SA2BSA) and Ab155) was performed with the support of the ICTS (Infraestructuras Científico-Tecnológicas Singulares “NANBIOSIS” Spain), more specifically by the Custom Antibody Service (CAbS, CIBER-BBN, IQAC-CSIC). The immunoreagents for sulfapyridine (SPy) detection used for the development of the biosensor were described before [23 (link)]. All other chemicals were purchased from Sigma-Aldrich and were used as received. All electrochemical measurements were performed on a VMP3 electrochemical workstation.