摘要
运用新型测定活细胞个数的化学物质,建立一种快速测定重组人白细胞介素-2生物学活性的方法.活细胞代谢过程中,胞内的可溶性噻唑盐WST-8在电子载体1-Methoxy PMS存在时被还原成可溶性甲臢染料,甲臢染料可在A45Onm处产生吸收值,根据其吸收值的大小间接检测活细胞个数从而测定待检测样品中IL-2的浓度水平.相关性实验结果证明,在细胞浓度1250~160000个/孔范围内,CTLL-2活细胞个数与A45O值之间呈线性相关;用此种新方法检测的白细胞介素-2(IL-2)生物学活性结果与MTT法及XTT法结果基本一致,相关系数分别为r=0.9602和r=0.9511.因此, WST-8法适用于悬浮依赖型细胞CTLL-2为基础的IL-2生物学活性检测,且具有经济、实用、简便、易行的特点,有较好的实际应用价值.
To develop a method for dermination of biological activity of recombinant human interleukin-2 with a new chemical substance WST-8. During the process of metabolism of living cells, in the presence of 1-Methoxy PMS, the WST-8 in cells may be reduced to soluble Formazon with the occurrence of absorbance at 450nm, which is dependent to the number of living cells. IL concentration in tested samples could be determined based on this deduction. The result showed that, at a certain range of cell concentration ,the number of living CTLL-2 cells was linearly related to A450. The IL-2 activity determined by WST-8 method was basically consistent with those by MTT ( r = 0. 954 ) and XTT ( r = 0. 955 ). WST-8 method was an economical and simple performance, and suitable for the determination of IL-2 activity based on IL-2 dependent CTLL-2.
出处
《微生物学免疫学进展》
2005年第4期31-34,共4页
Progress In Microbiology and Immunology