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Reagent and method for predicting curative effect of tumor necrosin inhibitor

A tumor necrosis factor and inhibitor technology, used in the field of reagents for predicting the efficacy of tumor necrosis factor inhibitors

Active Publication Date: 2018-03-20
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the study found that 40% of patients still did not respond to the drug

Method used

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  • Reagent and method for predicting curative effect of tumor necrosin inhibitor
  • Reagent and method for predicting curative effect of tumor necrosin inhibitor
  • Reagent and method for predicting curative effect of tumor necrosin inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0136] In this example, a total of 92 patient samples and clinical data were collected, including plasma samples obtained from 79 patients at 0, 2, 4, 8, and 12 weeks after administration to detect IgG galactose modification levels. Wherein the modification level of IgG galactose is carried out according to the method described in CN201510632081.1, including using mass spectrometry to determine the modification level of IgG galactose: G0 (no galactose modification); G1 (one galactose modification), G2 (two galactose modifications) , and quantitatively calculate the IgG galactose modification distribution (IgG-Gal ratio) based on the contents of these three galactose modifications. The specific steps are as follows:

[0137] 1. Purification of IgG from blood samples of screening subjects

[0138] Refer to the Thermo Fisher Scientific Protein A Spin Plate for IgG Screening user manual for implementation, as follows:

[0139] a. Equilibrate the purification column and buffer so...

Embodiment 2

[0168] Whole exome sequencing was performed on 24 responders and 9 nonresponders. Whole exome sequencing uses ultrasound to break up the genome sequence, then uses Roche's capture and library reagents, and then uses illumina Hiseq2000 sequencing. After obtaining the whole exome sequencing data, use Burrows Wheeler Alignment V.0.7.10 for sequence genome alignment, and genome analysis toolkit (GATK) V.1.1.28 for assembly and search for variant sites. Finally, Annovar was used to annotate the variant sites. According to the above process, six candidate gene-related loci were found, including MYOM2, VPS13B, DISP1 and IL27. By expanding the sample and using next-generation sequencing (Sanger) sequencing, it was found that the rs2294066 site of the MYOM2 gene (ie, the 322nd of SEQ ID NO: 3) was significantly different between responding patients and non-responding patients (OR=3.82, p value=1.1×10 -3 )(Table 1).

[0169] Table 1: Distribution of the MYOM2(rs2294066) locus between...

Embodiment 3

[0177] Since gene loci and IgG galactose modifications represent different genetic backgrounds and transcriptional post-translational modification levels of patients, the present invention predicts the curative effect of tumor necrosis factor antagonist drugs through the combination of the above two biomarkers, thereby assisting clinical more precise treatment.

[0178] according to image 3, when a patient with ankylosing spondylitis considers using tumor necrosis factor inhibitors (fusion protein drugs, such as Yisaipu), first measure the IgG galactose modification level and calculate the Gal ratio, if the Gal ratio is greater than 0.8, then It is predicted that the patient has a good response to the drug, and it is recommended to use the drug for treatment; if the Gal ratio is less than 0.8, it is recommended to use the drug once and measure the IgG galactose modification level again after 2 weeks to calculate the Gal ratio. If the Gal ratio measured after 2 weeks of medic...

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Abstract

The invention relates to a reagent and a method for predicting the curative effect of the tumor necrosin inhibitor. Particularly, the invention provides application of a reagent and / or an instrument to the preparation of a product for predicting the curative effects of treating patients with ankylosing spondylitis by using tumor necrosin inhibitors, wherein the reagent and / or instrument comprisesa tail end galactosylation level of the blood IgG surface double-antenna complicated N sugar chain, wherein the tail end galactosylation level is double-antenna complicated N-sugar chain level Ga10 connected with no galactose at the tail end, a double-antenna complicated N-sugar chain level Gal1 connected with one galactose at the tail end and a double-antenna complicated N-sugar chain level Gal2connected with two galactoses at the tail end on the IgG surface.

Description

technical field [0001] The field relates to reagents and methods for predicting the efficacy of tumor necrosis factor inhibitors. Background technique [0002] Ankylosing spondylitis is an immune-mediated inflammatory disease, which is mainly genetic and caused by a variety of infections. Tumor necrosis factor (TNF) is an extensively studied factor in ankylosing spondylitis. Multiple functional susceptibility loci of the tumor necrosis factor (TNF) family are associated with the pathogenesis of ankylosing spondylitis, and they participate in multiple pathogenic pathways. Therefore, tumor necrosis factor inhibitors are currently widely used to treat this disease. However, the study found that 40% of the patients did not respond to the drug. Therefore, it is urgent to find biomarkers to predict the curative effect of tumor necrosis factor in clinical practice. [0003] The glycosylation modification of protein can reflect the real intracellular environment. Immunoglobulin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N27/62G01N30/02G01N24/08
CPCG01N24/08G01N30/02G01N33/68G01N33/6848G01N33/6851
Inventor 王久存刘晶邹和建任士芳顾建新牛振民金力李媛马彦云杜爱平
Owner FUDAN UNIV
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