Inorganic salts, sucrose, Tween 80, human serum albumin, and gamma globulin (IgGHum) (AppliChem GmbH, Darmstadt, Germany and Sigma–Aldrich, St. Louis, MO, USA) were used in this work. A commercial set of bovine blood serum samples was used (Biolot, Saint Petersburg, Russia).
All experiments were performed in ultrapure water prepared with Milli-Q equipment (Merck KGaA, Darmstadt, Germany). PBS was prepared from tablets from EcoService (Saint Petersburg, Russia). HEPES buffer, pH 7.5, from AppliChem (Darmstadt, Germany), 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC-HCl) from Roth (Karlsruhe, Germany), and sodium salt of N-hydroxysulfosuccinimide (s-NHS) from Chem-Impex Int’l (Wood Dale, IL, USA) were used.
Xeomin 50 Units per vial (Merz therapeutics, Frankfurt am Main, Germany) contains BoNT serotype A in human serum albumin with sucrose. The powder was dissolved in 125 µL Ringer’s solution (Grotex, Saint Petersburg, Russia) providing a solution with 400 Units/mL BoNT, 8 mg/mL human serum albumin, and 38 mg/mL sucrose. The solution was exploited immediately after the preparation. According to Frevert [21 (link)], 400 Units/mL of Xeomin corresponds to 1.76 ng/mL or 11.7 pM of BoNT serotype A.
All experiments were performed in ultrapure water prepared with Milli-Q equipment (Merck KGaA, Darmstadt, Germany). PBS was prepared from tablets from EcoService (Saint Petersburg, Russia). HEPES buffer, pH 7.5, from AppliChem (Darmstadt, Germany), 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC-HCl) from Roth (Karlsruhe, Germany), and sodium salt of N-hydroxysulfosuccinimide (s-NHS) from Chem-Impex Int’l (Wood Dale, IL, USA) were used.
Xeomin 50 Units per vial (Merz therapeutics, Frankfurt am Main, Germany) contains BoNT serotype A in human serum albumin with sucrose. The powder was dissolved in 125 µL Ringer’s solution (Grotex, Saint Petersburg, Russia) providing a solution with 400 Units/mL BoNT, 8 mg/mL human serum albumin, and 38 mg/mL sucrose. The solution was exploited immediately after the preparation. According to Frevert [21 (link)], 400 Units/mL of Xeomin corresponds to 1.76 ng/mL or 11.7 pM of BoNT serotype A.
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