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Cell immobilization technology and AQP4 antibody detection kit prepared through same

A cell fixation and kit technology, applied in the medical field, can solve the problems of small number of cells, differences in transfection efficiency, cumbersome operation, etc., and achieve the effect of cheap manpower and material resources, short time and simple operation

Pending Publication Date: 2017-01-11
NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, this method is cumbersome to operate, the transfection efficiency of different wells in the same plate may be different, the number of fixed cells is small, and it is easy to fall off, and the sample volume required for detection is relatively large, so it is not suitable for clinical laboratory testing and testing. commercial production
Currently these kits can only be provided by Omen Medical Laboratory Diagnostics Co., Ltd.

Method used

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  • Cell immobilization technology and AQP4 antibody detection kit prepared through same
  • Cell immobilization technology and AQP4 antibody detection kit prepared through same
  • Cell immobilization technology and AQP4 antibody detection kit prepared through same

Examples

Experimental program
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Embodiment

[0059] 1. Preparation of AQP4 transfected cells

[0060] To retrieve the AQP4 protein gene and its sequence number from GeneBank, see the sequence table (NM_001650); the human AQP4 gene fragment was synthesized by "Nanjing GenScript Biotechnology Co., Ltd.", with restriction sites at both ends of the gene fragment being XhoI and Kpnl , And inserted into the pDsRed-Monomer-N1 plasmid, the plasmid vector is the finally constructed plasmid pDsRed-Monomer-N1-AQP4, the map is shown in the attachment figure 2 .Sequencing to verify the constructed plasmid, the plasmid sequence is shown in the sequence list (GenebankNM_001650, the base sequence between the start codon and the stop codon (64..1035)), and the sequencing result is shown Figure 3-5 .

[0061] 1.1 Use a petri dish with a diameter of 10cm to culture cells at a density of 2.2×10 6 Cells were inoculated in each / 10cm petri dish, the medium was complete medium (DMEN basic medium: fetal bovine serum: penicillin streptomycin mixture ...

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Abstract

The invention discloses a cell immobilization technology. The cell immobilization technology comprises the following steps that 1, a pDsRed-Monomer-AQP4 plasmid is constructed; 2, transfection is conducted on cells by the plasmid pDsRed-Monomer-AQP4 through a polyethyleneimine (PEI) transfection method; 3, the cells obtained after transfection are immobilized on a 96-well plate. According to the cell immobilization technology, a large quantity of AQP4 transfection cells can be effectively prepared at a time through the technology, then slide preparation is conducted, and therefore the cell immobilization technology is more suitable for commercialized production; meanwhile, in-batch difference caused by the transfection efficiency can be avoided, and the small size of a specimen needed by detection is needed. The invention further discloses an AQP4 antibody detection kit prepared through the technology.

Description

Technical field [0001] The invention relates to the medical field, in particular to a cell fixation process and the preparation of an AQP4 antibody detection kit through the process. Background technique [0002] Aquaporin 4 (AQP4) antibody, also known as neuromyelitis optica (NMO)-IgG antibody, AQP4 antibody is one of the important detection indicators in the NMO diagnosis and treatment guidelines. It is an important serum for distinguishing multiple sclerosis and NMO landmark. The elevation of AQP4 antibody in cerebrospinal fluid (abbreviated as CSF) also has important reference significance. [0003] Existing methods all adopt the overexpression of AQP4 protein in mammalian cells, and fix the overexpressed cells with the slide method or use specially treated glass slides to culture and fix the cells. However, this method is cumbersome to operate. The transfection efficiency of different wells in the same plate may be different, the number of fixed cells is small, and it is eas...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N11/00G01N33/68
CPCC12N15/85C12N11/00C12N2800/107G01N33/6854
Inventor 刘光辉谢作善潘速跃汪鸿浩胡亚芳顾勇许苗菁宋萍萍李伟
Owner NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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