Preparation and performance of recombinant human cystatin C quality control material

doi:10.3969/j.issn.1673-8640.2017.08.017

Abstract

Abstract:

Objective To prepare cystatin C（Cys C） quality control material by using recombinant human Cys C protein expressed in Escherichia coli BL21 as basic material. Methods By gene engineering method,recombinant human Cys C protein was expressed through pCold1 and BL21. Freeze drying was performed after purification. Results Inclusion bodies of recombinant human Cys C protein was expressed successfully. After renaturation by dialysis method,soluble recombinant human Cys C protein with immunological activity had been prepared. Cys C quality control material was prepared by using the recombinant human Cys C protein and determined by HITACHI and ROCHE analysis systems. Cys C quality control was suitable for the 2 analysis systems. Conclusions Cys C quality control material prepared by recombinant human Cys C protein has good stability and reliability,it could be used in various analysis platforms of in vitro diagnosis.

Key words: Cystatin C, Recombinant protein, Quality control material, Stability

CLC Number:

R446.1

LI Jiangfeng, FENG Minyi, QIAN Wei, ZHAO Xiaoni, CHEN Zitong, JIAO Liyuan, WANG Jihua, CAI Lei. Preparation and performance of recombinant human cystatin C quality control material[J]. Laboratory Medicine, 2017, 32(8): 733-737.

Figures/Tables 6

项目	第1批			第2批			第3批			第4批
项目	L1	L2	L3	L1	L2	L3	L1	L2	L3	L1	L2	L3
$x ?$ （mg/L）	0.72	2.27	6.27	0.66	2.23	6.82	0.63	2.25	6.53	0.77	2.68	6.88
CV（%）	3	1	2	4	3	4	3	2	3	2	1	2
s₁	0.026	0.035	0.099	0.026	0.077	0.292	0.028	0.040	0.185	0.022	0.034	0.134
s₂	0.008	0.011	0.029	0.009	0.022	0.019	0.016	0.019	0.030	0.013	0.023	0.040
上限（mg/L）	0.83	2.61	6.90	0.76	2.56	7.50	0.72	2.59	7.18	0.89	3.08	7.57
下限（mg/L）	0.61	1.93	6.21	0.56	1.90	6.75	0.54	1.91	6.46	0.65	2.28	6.19

项目	第1批			第2批			第3批			第4批
项目	L1	L2	L3	L1	L2	L3	L1	L2	L3	L1	L2	L3
$x ?$ （mg/L）	0.72	2.27	6.27	0.66	2.23	6.82	0.63	2.25	6.53	0.77	2.68	6.88
CV（%）	3	1	2	4	3	4	3	2	3	2	1	2
s₁	0.026	0.035	0.099	0.026	0.077	0.292	0.028	0.040	0.185	0.022	0.034	0.134
s₂	0.008	0.011	0.029	0.009	0.022	0.019	0.016	0.019	0.030	0.013	0.023	0.040
上限（mg/L）	0.83	2.61	6.90	0.76	2.56	7.50	0.72	2.59	7.18	0.89	3.08	7.57
下限（mg/L）	0.61	1.93	6.21	0.56	1.90	6.75	0.54	1.91	6.46	0.65	2.28	6.19

项目	第1批		第2批		第3批		第4批
项目	CYS5019	CYS5020	CYS5019	CYS5020	CYS5019	CYS5020	CYS5019	CYS5020
$x ?$ （mg/L）	0.71	3.19	0.59	3.41	0.65	3.38	0.62	3.27
s₁	0.022	0.017	0.025	0.027	0.021	0.031	0.019	0.016
s₂	0.011	0.012	0.008	0.008	0.019	0.021	0.008	0.008
CV（%）	3	0	4	1	1	1	3	0
上限（mg/L）	0.82	3.51	0.68	3.75	0.75	3.72	0.71	3.60
下限（mg/L）	0.60	2.87	0.50	3.07	0.55	3.04	0.53	2.94

项目	第1批		第2批		第3批		第4批
项目	CYS5019	CYS5020	CYS5019	CYS5020	CYS5019	CYS5020	CYS5019	CYS5020
$x ?$ （mg/L）	0.71	3.19	0.59	3.41	0.65	3.38	0.62	3.27
s₁	0.022	0.017	0.025	0.027	0.021	0.031	0.019	0.016
s₂	0.011	0.012	0.008	0.008	0.019	0.021	0.008	0.008
CV（%）	3	0	4	1	1	1	3	0
上限（mg/L）	0.82	3.51	0.68	3.75	0.75	3.72	0.71	3.60
下限（mg/L）	0.60	2.87	0.50	3.07	0.55	3.04	0.53	2.94

测试温度	测试天数（d）	低值（mg/L）	中值（mg/L）	高值（mg/L）
4 ℃	1	0.67	2.49	6.37
	2	0.70	2.41	6.34
	4	0.73	2.48	6.25
	8	0. 72	2.49	6.49
25 ℃	1	0.72	2.54	6.37
	2	0.72	2.60	6.35
	4	0.77	2.60	6.23
	8	0.76	2.65	6.86

测试天数（d）	低值（mg/L）	中值（mg/L）	高值（mg/L）
3	0.77	2.65	6.46
6	0.85	2.52	6.34
9	0.78	2.58	6.27
12	0.76	2.48	5.86

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