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CN-122017248-A - ELISA kit composition and kit for detecting advanced oxidized protein product

CN122017248ACN 122017248 ACN122017248 ACN 122017248ACN-122017248-A

Abstract

The invention belongs to the technical field of biology, and particularly relates to an ELISA kit composition and a kit for detecting a late oxidized protein product. The kit composition comprises the following raw materials of 20-50mM Tris-HCl, 100-200mM NaCl, 0.5% -2.0% w/v of fetal bovine serum, 0.1% -0.5% w/v of sodium caseinate, 1-5mM disodium ethylenediamine tetraacetate, 1% -5% w/v of trehalose, 0.05% -0.2% w/v of polyvinylpyrrolidone and 0.02% -0.05% v/v of ProClin 300. The kit composition provided by the invention is applied to a kit, so that the kit has better accuracy, sensitivity and stability.

Inventors

  • LI ZONGZE
  • ZHANG ZHONGMIN
  • XU JIAJIA

Assignees

  • 南方医科大学南方医院

Dates

Publication Date
20260512
Application Date
20260109

Claims (10)

  1. 1. An ELISA kit composition for detecting late oxidized protein products, comprising the following raw materials: 20-50mM Tris-HCl, 100-200mM NaCl, 0.5% -2.0% w/v fetal bovine serum, 0.1% -0.5% w/v sodium caseinate, 1-5mM disodium ethylenediamine tetraacetate, 1% -5% w/v trehalose, 0.05% -0.2% w/v polyvinylpyrrolidone and 0.02% -0.05% v/v ProClin.
  2. 2. The kit composition of claim 1, comprising the following materials: 30mM Tris-HCl, 150mM NaCl, 1.0% w/v foetal calf serum, 0.25% w/v sodium caseinate, 2mM disodium edetate, 3% w/v trehalose, 0.1% w/v polyvinylpyrrolidone and 0.03% v/v ProClin 300,300.
  3. 3. The kit of claim 1, wherein the pH of the kit is from 5.5 to 6.5.
  4. 4. The method for preparing the kit composition according to any one of claims 1 to 3, which is characterized by comprising the following steps of stirring and mixing Tris-HCl, naCl, fetal bovine serum, sodium caseinate, disodium ethylenediamine tetraacetate, trehalose and polyvinylpyrrolidone according to a proportion, adding ProClin to 300, mixing uniformly, fixing the volume, and filtering.
  5. 5. The method according to claim 4, wherein the filtration is performed with a 0.22 μm filter.
  6. 6. Use of a kit composition according to any one of claims 1-3 for the preparation of an ELISA kit for the detection of late oxidized protein products.
  7. 7. An ELISA kit for detecting late oxidized protein products comprising the kit composition of any of claims 1-3.
  8. 8. The ELISA kit of claim 7, further comprising a quality control and a calibrator.
  9. 9. The ELISA kit according to claim 7, wherein the calibrator is prepared by dissolving the late oxidized protein product as a raw material of the calibrator in the kit composition according to any one of claims 1 to 3, and mixing the solution thoroughly.
  10. 10. The ELISA kit according to claim 7, wherein the quality control product is prepared by dissolving the advanced oxidized protein product as a raw material of the quality control product with the kit composition according to any one of claims 1 to 3, and mixing the dissolved and dissolved product.

Description

ELISA kit composition and kit for detecting advanced oxidized protein product Technical Field The invention belongs to the technical field of biology, and particularly relates to an ELISA kit composition and a kit for detecting a late oxidized protein product. Background The late oxidized protein product (AOPPs) is a protein cross-link containing bistyrosine formed by proteins, particularly albumin, in myeloperoxidase-mediated oxidation reactions. The elevated levels are closely related to the severity and prognosis of end stage renal disease, diabetic complications, inflammatory disease, and cardiovascular disease. The enzyme-linked immunosorbent assay has become the mainstream technology for detecting AOPPs due to its high throughput, relatively simple operation and good specificity. The AOPP ELISA kit on the market at present mostly adopts the principle of a double antibody sandwich method or a competition method, but most of the AOPP ELISA kits have the problems of insufficient sensitivity and specificity, poor long-term stability, weak anti-interference capability and the like, and restrict the detection accuracy and application breadth. For example, patent CN116375855A discloses a monoclonal antibody specifically recognizing AOPPs and its application. The inventor obtains hybridoma cell strain (preservation number CCTCCNO. C202232) secreting monoclonal antibody AP-4C5 through experimental screening, and monoclonal antibody AP-4C5 can specifically recognize an oxidation group in AOPPs and does not react with natural protein. A sandwich ELISA system is established by using a single AP-4C5, using the AP 4C5 as a coating antibody and using HRP-AP-4C5 as a detection antibody, the concentration of 0-20 mu g/ml of AOPP-HSA and AOPP-FB can be detected, and AOPPs in human serum can be quantitatively detected, and using a commercial anti-human albumin (HSA) polyclonal antibody as a coating antibody and using HRP-AP-4C5 as a detection antibody, a multi-monoclonal antibody sandwich ELISA system is established, the concentration of 0-40 mu g/ml of AOPPs can be detected, and AOPPs in the plasma of healthy people and patients can be quantitatively detected. However, the long-term stability of the invention is still to be improved. Such as literature, "research on the molecular mechanism of the induction of chondrocyte inflammatory response by advanced oxidized protein products", university of southern medical science, 2019.000600. This study demonstrates the effect of late oxidized protein products on interleukin and tumor necrosis factor expression. However, the study was not optimal for the kit related to the advanced oxidized protein product. Therefore, there is a need to study an ELISA kit composition for detecting advanced oxidized protein products, which has high sensitivity and specificity, good long-term stability and high detection accuracy. Disclosure of Invention The invention aims to provide an ELISA kit composition and a kit for detecting a late oxidized protein product, and a related technology thereof, so as to solve the technical problems of insufficient sensitivity and specificity, poor long-term stability, low accuracy and the like of the ELISA kit for detecting the late oxidized protein product in the prior art or the combination thereof. Description of the terminology: Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs. All patents, patent inventions, and publications cited herein are incorporated by reference in their entirety unless otherwise indicated. If there are multiple definitions of terms herein, the definitions of this chapter shall control. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the subject matter of the invention, as claimed. In the present invention, the singular is used to include the plural unless specifically stated otherwise. It should also be noted that the use of "or" means "and/or" unless stated otherwise. Furthermore, the terms "include," as well as other forms, such as "comprising," "including," and "containing," are not limiting. Unless specifically defined, the use of various commercially available products as used herein employs standard techniques. For example, the kit may be used by the manufacturer, or may be implemented in a manner known in the art or in accordance with the description of the invention. The techniques and methods described above may generally be practiced according to conventional methods well known in the art, based on a number of general and more specific descriptions in the literature cited and discussed in this specification. In a first aspect, the invention provides an ELISA kit composition for detecting late oxidized protein products, comprising the following raw materials: