Double-antibody sandwich chemiluminescence immune assay method

A chemiluminescence immunoassay and double-antibody sandwich technology, applied in the field of biomedicine, can solve the problems of low sensitivity, inability to provide the range of serum RPTN content, and lack of quality control indicators for clinical testing reagents.

Inactive Publication Date: 2018-08-10
SHAANXI NORMAL UNIV
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Problems solved by technology

The existing RPTN enzyme-linked analysis method is only used for scientific research, and the sensitivity is too low, and it also lacks the quality control indicators required for clinical test reagents, and cannot provide the content range of serum RPTN in normal people and patients with schizophrenia

Method used

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Embodiment Construction

[0012] The present invention will be further described below in conjunction with embodiment.

[0013] Analyze the epitope of the human RPTN protein sequence, design the N-terminal epitope sequence, and design the C-terminal epitope sequence; after cross-linking the N-terminal epitope sequence with KLH protein, immunize Balb mice, and prepare spleen Cells, and fused with myeloma cells, and then prepare N-terminal monoclonal antibody, coated on the microtiter plate, as a capture antibody; after cross-linking the C-terminal epitope sequence with KLH protein, immunize New Zealand rabbits to prepare anti- C-terminal rabbit polyclonal antibody, anti-C-terminal rabbit polyclonal antibody is used as detection antibody; anti-RPTN N-terminal mouse monoclonal antibody is used as capture antibody to coat on the microtiter plate, after washing, add the serum to be tested or RPTN standard, after washing Add anti-RPTN C-terminal rabbit polyclonal antibody as detection antibody; add goat anti...

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Abstract

The invention belongs to biomedical field, and relates to a double-antibody sandwich chemiluminescence immune assay method. The method comprises the following steps: adopting an anti-RPTN N-terminal mouse monoclonal antibody as a capturing antibody coated on an elisa plate, adding serum to be measured or RPTN standard after washing, and adding an RPTN C-terminal rabbit polyclonal antibody as a detection antibody after washing; adding goat anti-rabbit IgG-HRP after washing, and adding a luminal reaction solution after washing; and measuring chemiluminiscence (RLU) on a chemiluminiscence immuneanalyzer after washing, wherein the intensity of RLU represents the content of RPTN. The method firstly discovers the phenomenon that the RPTN inside the serum of a schizophrenic patient is reduced, an RPTN chemiluminescence immune method is further developed and the normal range of the RPTN inside the serum is detected and determined, and a novel method is provided for diagnosis and therapeutic effect evaluation of schizophrenia; and a novel tool is provided for the research of other neuropsychiatric diseases and the research and development of antipsychotic drugs.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to a double-antibody sandwich chemiluminescent immunoassay method. Background technique [0002] Currently, methods such as the Positive and Negative Syndrome Scale (PANSS) are mostly used in the diagnosis of schizophrenia and the evaluation of drug efficacy, but the research on serological indicators still has no ideal markers). RPTN is a member of the highly conserved S100 calcium-binding protein family in mammals, and was previously considered to be a protein unique to skin cells involved in the formation of the skin barrier. We reported that patients with schizophrenia and chronic methamphetamine (meth) users had significantly lower serum RPTN levels than normal individuals ( figure 1 , a), and chronic methamphetamine users are considered as a model of human schizophrenia; at the same time, we found that the atypical anti-schizophrenic drug Olanzapine (Olanzapine) can significantly i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 赵利军
Owner SHAANXI NORMAL UNIV
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