Preparation method of latex microsphere cross-linking specific monoclonal or polyclonal antibody for detecting soluble abnormally glycosylated modified CD58 molecule

A polyclonal antibody, glycosylation technology, applied in the field of medical testing, can solve the problems of long detection time and low sensitivity

Inactive Publication Date: 2017-03-22
ZHEJIANG UNIV
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Problems solved by technology

The traditional enzyme-linked immunosorbent assay is gradually replaced by new technology due to its long detection time and low sensitivity

Method used

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  • Preparation method of latex microsphere cross-linking specific monoclonal or polyclonal antibody for detecting soluble abnormally glycosylated modified CD58 molecule
  • Preparation method of latex microsphere cross-linking specific monoclonal or polyclonal antibody for detecting soluble abnormally glycosylated modified CD58 molecule

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Embodiment Construction

[0024] In order to make the technical means, creative features, goals and effects achieved by the present invention easy to understand, the present invention is further described using examples.

[0025] 1. Preparation method of abnormally glycosylated CD58 molecule monoclonal IgG antibody

[0026] 1.1 Animal immunity

[0027] The total RNA of 1х106 colorectal cancer LoVo cells was isolated and purified with RNAeasy kit, the total RNA was reverse transcribed into cDNA by Oligo dT15 primer, the human CD58 gene sequence (NM_001779.2) was amplified by polymerase chain reaction (PCR), and analyzed by EcoRI After digestion with / BamH1, it was cloned into the pLXSN retroviral vector, verified by DNA sequencing, a CD58 expression retroviral vector system was constructed, transfected into PT67 cells to produce recombinant retroviruses, and infected with colorectal cancer SW620 cells at a titer of 10:1. Positive clones were screened for 7-14 days with 500-700 μg / mL of G418 drug pressu...

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Abstract

The invention relates to a preparing method of a latex microsphere crosslinked specific monoclonal or polyclonal antibody for detecting soluble abnormally glycosylated CD58 molecules. The preparing method includes steps of 1) activating latex microspheres, 2) crosslinking the latex microspheres and anti-CD58 monoclonal and polyclonal antibodies, 3) washing the crosslinked compound, and storing the crosslinked compound. The latex microsphere crosslinked specific monoclonal or polyclonal antibody prepared by the method can be subjected to specific bonding reactions with abnormally glycosylated CD58 protein, while non-specific reactions don't happen between the monoclonal or polyclonal antibody and normally expressed CD58 protein and non-CD58 protein. The latex microsphere crosslinked specific monoclonal or polyclonal antibody is used for detecting the abnormally glycosylated CD58 molecular level of peripheral blood or serums in latex immunoturbidimetry, and used for quantitative analysis of the content of abnormally glycosylated CD58 in samples.

Description

technical field [0001] This patent relates to the field of medical testing, in particular to a method for preparing latex microsphere cross-linking specific monoclonal or polyclonal antibodies for detecting soluble abnormally glycosylated modified CD58 molecules. Background technique [0002] CD58 molecule is lymphocyte function associated antigen-3 (lymphocyte function associated antigen-3, LFA-3), is a transmembrane protein molecule on the cell surface, belongs to the cell adhesion molecule of the immunoglobulin superfamily, and is expressed in hematopoietic cells. Vascular endothelium and epithelial cells. The CD58 molecule is the ligand of the CD2 molecule on the surface of T cells and natural killer cells (NK cells). In vitro studies have shown that the combination of CD2 and CD58 can cause the proliferation and activation of lymphocytes. This effect does not require the participation of auxiliary cells such as macrophages. Antigen non-specific, as a bypass system for ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/531G01N33/68G01N33/574
CPCG01N33/531G01N33/57484G01N33/6803
Inventor 朱永良秦光明吴佳
Owner ZHEJIANG UNIV
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