Establishment and validation of a time-resolved fluoroimmunoassay for transient receptor potential channel 5

doi:10.3969/j.issn.1673-8640.2018.05.017

Abstract

Abstract:

Objective Transient receptor potential channel 5（TRPC5） can be as a marker of breast cancer drug resistance. To establish a method for the diagnosis of breast cancer in vitro drug resistance. Methods Time-resolved fluoroimmunoassay（TRFIA）was established for TRPC5. Anti-TRPC5 antibody was prepared by immunizing New Zealand white rabbits with TRPC5-keyhole limpet hemocyanin （KLH） as an immunogen. Mucin 1 （MUC1） antibody coated immunomagnetic beads to process samples. TRPC5 peptides, coated with 96-well plates as solid-phase antigens, competed with free TRPC5 antigen. Eu³⁺-goat anti-rabbit antibody was used as a tracer. Results The sensitivity was 1 ng/mL for TRPC5 TRFIA. The range was 5-500 ng/mL. The coefficient of variation（CV）was 5.5%. The mean recovery rate was 94.6%. The ED80,ED50 and ED20 of TRFIA were（4.68±0.13）,（321.66±2.77） and （714.42±4.07）ng/mL,respectively. There was statistical significance for the results of chemotherapy-resistant patients and chemotherapy-tolerant patients（P<0.000 1）. Conclusions TRPC5 TRFIA has a certain degree of differentiation between breast cancer chemotherapy resistance or not. It has clinical significance and is worth further study.

Key words: Transient receptor potential channel 5, Time-resolved fluoroimmunoassay, Breast cancer, Chemotherapy resistance

CLC Number:

R446.61

LU Yangfan, HUANG Biao, MA Xin. Establishment and validation of a time-resolved fluoroimmunoassay for transient receptor potential channel 5[J]. Laboratory Medicine, 2018, 33(5): 447-451.

Figures/Tables 8

References 11

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组别	1	2	3	4	5	6	x	CV（%）
1	1 047 562	1 028 169	1 121 664	1 135 837	1 056 903	1 005 637	1 065 962	4.9
2	1 163 358	1 151 077	1 177 126	1 120 490	1 124 884	1 240 175	1 162 851	3.8
3	1 121 710	1 106 639	1 105 110	1 072 046	1 058 466	1 147 603	1 101 929	3.0
4	1 084 164	1 182 513	1 183 555	1 203 036	1 183 232	1 214 754	1 175 209	4.1
x				1 126 488
CV（%）				5.5

组别	1	2	3	4	5	6	x	CV（%）
1	1 047 562	1 028 169	1 121 664	1 135 837	1 056 903	1 005 637	1 065 962	4.9
2	1 163 358	1 151 077	1 177 126	1 120 490	1 124 884	1 240 175	1 162 851	3.8
3	1 121 710	1 106 639	1 105 110	1 072 046	1 058 466	1 147 603	1 101 929	3.0
4	1 084 164	1 182 513	1 183 555	1 203 036	1 183 232	1 214 754	1 175 209	4.1
x				1 126 488
CV（%）				5.5

样本	TRPC5 添加量（ng/mL）	TRPC5 TRFIA测量值（ng/mL）	TRPC5 TRFIA测定回收率（%）
1	10	11.01	110.12
2	50	47.28	94.57
3	100	85.51	85.51
4	500	466.48	93.30
5	800	716.57	89.57

样本	TRPC5 添加量（ng/mL）	TRPC5 TRFIA测量值（ng/mL）	TRPC5 TRFIA测定回收率（%）
1	10	11.01	110.12
2	50	47.28	94.57
3	100	85.51	85.51
4	500	466.48	93.30
5	800	716.57	89.57

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