阅读说明：本技术 催化生物陶材料从含目的蛋白的样品中去除病毒的方法 (Method for removing viruses from target protein-containing sample by catalyzing bioceramic material ) 是由 汪涛 严娜 曹忆 周洁 殷梦琪 钱星 孙莉洁 于 2020-08-04 设计创作，主要内容包括：本发明公开了一种催化生物陶材料从含目的蛋白的样品中去除病毒的方法,利用多孔陶瓷颗粒制备成的正电荷多孔陶瓷颗粒可以在层析过程中特异性的吸附含病毒单克隆抗体溶液中的带负电荷的病毒颗粒,去除效果达到了6logs以上,而且单克隆抗体溶液的蛋白浓度未降低,达到了很好的病毒去除效果,比现有的先经过阴离子层析或亲和层析再加入纳滤去除病毒的方法,节约时间及纳滤耗材,并且蛋白溶液在层析设备及纳滤设备上总体时间大大减少,蛋白溶液的活性降低及总量增加。(The invention discloses a method for removing viruses from a sample containing target proteins by catalyzing a bioceramic material, wherein positively charged porous ceramic particles prepared by utilizing the porous ceramic particles can specifically adsorb virus particles with negative charges in a virus-containing monoclonal antibody solution in a chromatography process, the removal effect reaches over 6logs, the protein concentration of the monoclonal antibody solution is not reduced, and a good virus removal effect is achieved.)

1. A method of catalyzing the removal of a virus from a sample containing a protein of interest from a bioceramic material, comprising:

pretreating diatomite porous ceramic particles and then drying at room temperature;

soaking the dried diatomite porous ceramic particles in NaOH and YCl₃·6H₂In the sol solution prepared by O, ultrasonic coating reaction is carried out for 0.5 hour, and the sol solution is dried in a drying oven at 100 ℃, wherein Y is yttrium;

repeating the operation for three times, then placing the ceramic particles in a muffle furnace for dry baking at 700 ℃ and preserving heat for 1 hour to prepare positive charge porous ceramic particles;

screening positive charge porous ceramic particles through a screen;

loading the screened positive charge porous ceramic particles into a column in a hollow chromatographic column;

preparing a monoclonal antibody sample simulation solution, wherein the monoclonal antibody sample simulation solution comprises a bovine serum albumin standard solution and a virus, the ratio of the bovine serum albumin standard solution to the virus is 500:1, and the virus is a murine parvovirus or reovirus type 3;

enabling the monoclonal antibody sample simulation solution to flow through a chromatographic column taking positive charge porous ceramic particles as fillers;

collecting the flow-through liquid, the flushing liquid, the eluent and the cleaning liquid, and respectively determining the titer in the flow-through liquid, the flushing liquid, the eluent and the cleaning liquid.

2. The method for catalyzing removal of viruses from a target protein-containing sample according to claim 1, wherein the screened positively charged porous ceramic particles are loaded into a column in which the diameter of the column is 1cm to 1.5cm and the packing density of the particle basis weight is 0.5 kg/l.

3. The method for catalyzing a bioceramic material to remove viruses from a target protein-containing sample as recited in claim 1, wherein the step of loading the screened positively charged porous ceramic particles into a hollow chromatography column comprises:

adding 10ml of settled positive charge porous ceramic particles into an empty chromatographic column;

rinsing with 50ml of deionized water was carried out to equilibrium.

4. The method of catalyzing removal of a virus from a sample comprising a protein of interest according to claim 3, wherein after rinsing to equilibrium with 50ml of deionized water, the method further comprises:

the packed column was washed with 50ml of 0.5M NaOH solution at a rate of 1.0 ml/min.

5. The method for catalyzing removal of viruses from a sample containing target proteins of claim 4, wherein after washing the packed column with 50ml of 0.5M NaOH solution at a rate of 1.0ml/min, the method further comprises:

and (3) balancing the filled chromatographic column by using 120ml of balance buffer solution at the flow rate of 2.0ml/min, and setting the ultraviolet absorbance value to zero when the baseline of the absorbance value at the ultraviolet 280nm wavelength is stable.