分别依次使用1M NaOH 溶液和1M NaCl 溶液冲洗20 CV；然后用平衡缓冲液平衡50 CV；按10 CV/min的速度上样，收集UV-280处的吸收峰，上样完成后，相同的流速下使用平衡缓冲液继续冲洗，直到UV-280吸收值到基线，同时收集UV-280处吸收峰。
Flushing 1 Mol NaOH solution and 1 Mol NaCl solution 20CV seperately; With buffer liquid equilibrium 50 CV and on sample in the speed of 10 CV/min,collecting the absorption peak of UV-280,after sampling, using buffer liquid continue on flushing with same speed until the UV-280 absorption value to baseline and collecting the absoption peak of UV-280 as well.
在150L流加工艺建立中，工作体积XXL，接种密度XX/ml，控制Temp XX℃，DO XX%，Agit前期XXrpm，pH前期XX，后期XX，Gas前期XX SLPM。当活细胞密度大于XX/ml，进行第一次流加；当活细胞密度大于XX/ml，进行第二次流加；在第二次流加培养2天后，进行第三次流加，当细胞活性降至XX%，结束培养。
During the Fed-batch process of 150L,the volume of work is XXL,Inoculaton density is XX/ml,controlling the temperaure to XX℃,DO XX %,Agit XXrpm on earlier stage,PH XX on earlier stage and XX on later period,Gas is XX SLPM on earlier stage . When the viable cell density is geater than XX/ml we’ll start the first Fed-batch process, When the viable cell density is geater than XX/ml then begin the second Fed-batch process next after two days of cultivation,we will continue the third Fed-batch process, the whole cultivation will be end up with the cell competency down to XX%.
 
为了确证表达产物一级结构的正确性，企业自行建立了高效液相色谱与质谱联用肽图技术：先将XXXX蛋白经过变性、还原和烷基化，再通过胰蛋白酶将其切成不连续的多肽片段，然后利用高效液相色谱将酶切得到的所有肽段进行分离，再连接进入质谱分析每个肽段的准确分子量，通过二级质谱确认各肽段的氨基酸序列。综合所有鉴定出的肽段序列后与理论蛋白全序列比对，从而计算出蛋白总氨基酸序列覆盖率。
In order to express the correctness of the product primary structure,enterprise build the HPIC and peptide mapping technology on their own: first denaturate,return and alkylate the xxxx protein,then cut it into disconnected peptide fragment by trypsase,next separate all the atriopetion which cut with HPIC enzyme, final connect all the accurate molecular weight of each atriopetion which join in mass spectrum analysis  and confirm the amino acid sequence of each atriopetion by secondary mass spectrum.  After comparing all the confirmed atriopetion and theory of protein sequence then calculate the coverage rate of otal amino acid sequence of proteins.

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