Rituximab/graphene oxide composite antibody as well as preparation method and application thereof
A composite technology of rituximab and graphene, which is applied in the direction of antibodies, pharmaceutical formulations, anti-tumor drugs, etc., can solve the problem of weak curative effect of rituximab and achieve the effect of enhancing biological response
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[0038] Another aspect of the present invention provides a method for preparing a rituximab / graphene oxide composite antibody, comprising the following steps: adding rituximab and graphene oxide to a phosphate buffer with a volume percentage of 10-50% solution, shake well at 37-42°C, and react for 4-24 hours to obtain the rituximab / graphene oxide composite antibody.
[0039] Phosphate buffer saline (PBS) is a commonly used reagent in this field, and its pH range is 7.35-7.45. The volume percentage is 10-50% phosphate buffer saline solution, that is, PBS solution, and the addition of water is adjusted on the basis of the original formula to dilute it.
[0040] Rituximab and graphene oxide react under the condition of shaking and shaking, and the two are combined by non-covalent bond to obtain rituximab / graphene oxide composite antibody. And the rituximab / graphene oxide composite antibody can be separated from the phosphate buffer as a solvent, or it can be kept in the phosphate...
Embodiment 1
[0054] Graphene oxide raw materials were purchased from Sigma, purified by dialysis to remove alcohol, and prepared to a concentration of 1mg / ml. Under the condition of Misonix ultrasonic breaker Sonicator 3000 intensity 3.5 in the United States, the ultrasound was continued for 2 hours, the mode worked for 10 seconds, and rested for 5 seconds. Note that the whole process is carried out in a mixture of ice and water. After sonication, filter with a 0.22 μm filter to prepare graphene oxide, measure the final concentration with Nanodrop, and refrigerate for future use.
[0055] Add 1 mg of rituximab and 0.2 mg of graphene oxide into 1 ml of 10% phosphate buffer by volume, shake well at 4°C, and react for 12 hours to obtain rituximab / graphene oxide containing Mixture of complex antibodies.
Embodiment 2
[0057] The difference from Example 1 is that the temperature for preparing the rituximab / graphene oxide composite antibody is 37°C.
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