▎ 摘　　要

NOVELTY - Electrochemiluminescence (ECL) sensor is prepared by producing graphene oxide by Hummers method which involves mixing 1 g graphite, 1 g sodium nitrate (NaNO3), and 46 ml 98% sulfuric acid (H2SO4) under ice bath condition, adding slowly 6 g potassium permanganate (KMnO4), stirring at 35 degrees C for 1 hour, adding 80 ml pure water, stirring for 30 minutes, adding 200 ml water, adding slowly 6 ml 30% hydrogen peroxide (H2O2), reacting at room temperature for 1 hour, filtering, washing filtrate with water until pH is neutral, dispersing in 500 ml pure water for 2 hours to obtain dispersed GO. USE - Preparation of electrochemiluminescence (ECL) sensor for detecting kinase protein activity (claimed). ADVANTAGE - The sensor has high sensitivity, low detection limit, and good stability. DETAILED DESCRIPTION - Electrochemiluminescence resonance energy transfer (ECL) is prepared by producing graphene oxide by Hummers method which involves mixing 1 g graphite, 1 g sodium nitrate (NaNO3), and 46 ml 98% sulfuric acid (H2SO4) under ice bath condition, adding slowly 6 g potassium permanganate (KMnO4), stirring at 35 degrees C for 1 hour, adding 80 ml pure water, stirring continuously for 30 minutes, adding 200 ml water, adding slowly 6 ml 30% hydrogen peroxide (H2O2), reacting at room temperature for 1 hour, filtering, washing filtrate with water until pH is neutral, dispersing in 500 ml pure water for 2 hours to obtain dispersed graphene oxide (GO), preparing graphene quantum dots (GQDs) by hydrothermal method which involves drying GO in tubular furnace under nitrogen protection, heating at 200 degrees C for 2 hours and 5 degrees C/min heating rate to obtain graphene sheet, immersing 0.05 g graphene sheet in concentrated sulfuric and nitric acid mixed solution at volume ratio concentrated sulfuric acid:concentrated nitric acid of 1:3 for 17 hours, diluting with 250 ml water, filtering using 0.22 mu m micro-porous filtering film, collecting suspension, adding 40 ml water, adjusting pH to 8 by adding sodium hydroxide (NaOH) solution, putting in reaction container at 200 degrees C, reacting for 12 hours, cooling to room temperature, filtering using 0.22 mu m micro-porous filtering film to obtain GQD brown filtrate, mixing 0.05 g sodium hydroxide, and 0.1 g sodium acetate in 20 ml GQD solution, reacting for 3 hours, adding hydrochloric acid to neutralize and obtain carboxylated GQD, preparing anti-phosphoserine antibody graphene composite by mixing 200 mu l 1 mg/ml GO and 200 mu l 5 mg/ml anti-phosphoserine antibody, reacting at room temperature for 12 hours, centrifuging at 10000 rpm for 30 minutes, washing precipitate with ultrapure water for 3 times to obtain 10 mM phosphate buffer solution with 7.4 pH, maintaining temperature at 40 degrees C, preparing electrochemiluminescence (ECL) sensor by taking glassy carbon electrode with 1, 0.3, and 0.05 mu m particle sizes, polishing with alpha -aluminum oxide ( alpha -Al2O3) polishing paste, performing ultrasonic cleaning with ethanol and water for 1 minute, coating glassy carbon electrode with 10 mu l 0.5% chitosan solution, drying, immersing in solution containing 5 mM N-ethyl-N'-1-(3-dimethylaminopropyl) carboxylated GQD carbodiimide hydrochloride solution, incubating at room temperature for 5 hours, washing electrode with buffer solution, immersing in solution containing 5 mM N-ethyl-N'-1-(3-dimethylaminopropyl) carbodiimide hydrochloride, 8 mM N-hydroxysuccinimide, and 50 mu m polypeptide solution, reacting for 3 hours, immersing electrode in solution containing tris (hydroxymethyl) aminomethane hydrochloride buffer, protein kinase, and adenosine triphosphate, reacting for 2 hours to obtain phosphorylated polypeptide, and incubating for 1 hour. An INDEPENDENT CLAIM is included for detecting kinase protein activity which involves increasing concentration of kinase protein to 0.01-5 U/ml, performing phosphorylation to obtain modified electrode surface polypeptide, assembling GQDs and GO in modified electrode surface by immersing in phosphate buffer solution containing 0.1 M sodium persulfate (Na2S2O8) and 0.1 M potassium chloride (KCl), strengthening assembly by quenching, reducing ECL signal, detecting at 0.023 U/ml detection limit to obtain 0.15 ellagic acid, increasing ECL sensor to maximum, and calculating 0.043 mu M ellagic acid concentration.