Establishment of an ELISA for the quantitative measuring of glucagon-like peptide 1 and the primary clinical application

doi:10.3969/j.issn.1673-8640.2015.08.015

Abstract

Abstract:

Objective To establish an enzyme-linked immunosorbent assay (ELISA) for the quantitative measuring of glucagon-like peptide 1(GLP-1) in serum and to investigate the application in the individualized treatment of type 2 diabetes mellitus (T2DM) patients. Methods Using a polyclonal antibody as capture antibody, another monoclonal antibody as labeled antibody, serial dilutions as standard material, an ELISA was established for the quantitative measuring of GLP-1 in serum, and the biological reference interval was also established. All the subjects were classified into 3 groups, normal glucose tolerance (NGT) group (50 cases), impaired glucose tolerance (IGT) group (52 cases) and T2DM group (68 cases). The blood samples were collected at fasting time, 1 h after taking glucose and 2 h after taking glucose. The dynamic change of GLP-1 was evaluated with routine biological items, in order to evaluate the clinical application significance of GLP-1 for the treatment of T2DM. Results The established method had an good quantitative correlation in the range of 23.5-1 490 nmol/L(R²=0.999, P<0.05). The biological reference interval for adults was (79.5±15.6) nmol/L. The GLP-1 in T2DM group was higher than those in NGT and IGT groups (P<0.05). According to the dynamic change of GLP-1 concentration in the oral glucose tolerance test, the deficient secretion of GLP-1 after taking glucose among patients in IGT and T2DM groups could be concluded. Conclusions The ELISA for the quantitative measuring of GLP-1 in serum is successfully established. This method could be used to dynamically monitor the secretion of GLP-1 in IGT and T2DM patients and would be helpful for the individualized treatment of T2DM.

Key words: Glucagon-like peptide 1, Enzyme-linked immunosorbent assay, Type 2 diabetes mellitus, Impaired glucose tolerance

CLC Number:

R446.61

SUN Lishan, WANG Mingdong, LU Liu, LIU Yingbing, LU Lin, GUO Jinhu, FAN Lieying. Establishment of an ELISA for the quantitative measuring of glucagon-like peptide 1 and the primary clinical application[J]. Laboratory Medicine, 2015, 30(8): 835-840.

Figures/Tables 6

项目	GLP-1标准液浓度(nmol/L)
项目	1 490	745	373	186	93	47	23.5	0
$x ?$ (n=10)	2.831	1.662	0.971	0.580	0.347	0.204	0.088	0.029
SD	0.124	0.104	0.078	0.045	0.027	0.022	0.015	0.009
99% 可信区间	2.459~3.023	1.351~1.973	0.736~1.206	0.444~0.710	0.264~0.429	0.136~0.271	0.043~0.133	0.002~0.056
CV(%)	4.38	6.24	8.07	7.76	7.81	11.02	17.00	31.50

项目	GLP-1标准液浓度(nmol/L)
项目	1 490	745	373	186	93	47	23.5	0
$x ?$ (n=10)	2.831	1.662	0.971	0.580	0.347	0.204	0.088	0.029
SD	0.124	0.104	0.078	0.045	0.027	0.022	0.015	0.009
99% 可信区间	2.459~3.023	1.351~1.973	0.736~1.206	0.444~0.710	0.264~0.429	0.136~0.271	0.043~0.133	0.002~0.056
CV(%)	4.38	6.24	8.07	7.76	7.81	11.02	17.00	31.50

样本	检测次数	血清总GLP-1(nmol/L)		CV (%)
样本	检测次数	批内	批间	批内	批间
健康人	20	89.2±4.3	89.5±6.4	4.8	7.1
T2DM患者	20	201.0±5.5	201.5±13.7	3.7	6.8

组别	年龄(岁)	例数(男/女)	GLU(mmol/L)	IN (IU/mL)	GLP-1(nmol/L)	TG(mmol/L)
NGT组	45.3±14.9	50(28/22)	4.7±0.4	10.3±5.9	80.0±19.7	1.1±0.4
IGT组	53.7±13.2	52(29/23)	5.9±1.1	15.3±21.7	110.5±16.4	1.6±0.3
T2DM组	58.5±16.8	68(37/31)	8.2±2.5^*	26.7±14.3^*	145.1±24.5^*	1.4±0.2
组别	CHOL(mmol/L)	LDL-C(mmol/L)	HDL-C(mmol/L)	HbA_1c(%)	hs-CRP(mg/L)
NGT组	4.5±2.4	2.5±1.6	1.80±0.60	4.1±1.2	2.8±1.5
IGT组	4.6±0.3	3.2±0.3	1.21±0.14	6.2±0.2^*	13.7±3.2^*
T2DM组	4.2±0.3	2.8±0.2	1.14±0.07	7.0±0.4^*	13.6±3.4^*

组别	例数	GLP-1(nmol/L)
组别	例数	空腹	服糖后1 h	服糖后2 h
NGT组	50	80.0±19.7	150.0±67.9^#	70.4±39.8
IGT组	52	110.5±16.4	65.9±12.9^**##	69.5±27.1^##
T2DM组	68	145.1±61.3^**	104.2±42.1^*#	125.4±41.4^**

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