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Fluorescent immunochromatographic test paper for detecting human pgii protein and preparation method thereof

A fluorescence immunochromatography, test paper technology, applied in biological testing, measuring devices, analytical materials, etc., to achieve the effect of simple operation, good sensitivity, suitable for large-scale production

Active Publication Date: 2016-07-06
朱建安
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently no fluorescent immunochromatographic test strip for human PGII protein

Method used

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  • Fluorescent immunochromatographic test paper for detecting human pgii protein and preparation method thereof
  • Fluorescent immunochromatographic test paper for detecting human pgii protein and preparation method thereof
  • Fluorescent immunochromatographic test paper for detecting human pgii protein and preparation method thereof

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preparation example Construction

[0054] The preparation method of the PGII antigenic epitope peptide of the present invention can be chemical synthesis method: the antigenic epitope peptide is synthesized by a solid-phase method using an American ABI431A type polypeptide automatic synthesizer. The molecular weights of the antigenic epitope peptides (1) and (2) of the present invention are 1299.66 and 1943.41 respectively, which can be determined by mass spectrometry, and the synthesized antigenic epitope peptide sequences are identified by polypeptide sequence determination. The purity of the peptides was evaluated by thin-layer chromatography and high-performance liquid chromatography, and the concentration of the epitope peptide was determined.

[0055] 2. PGII antigen

[0056] The present invention also provides a PGII antigen prepared by coupling one of the human PGII antigen epitope peptides (1) and (2) of the present invention with a carrier protein. Specifically, the present invention provides PGII an...

Embodiment 1

[0107] Example 1: Preparation of PGII epitope peptides (1) and (2).

[0108] The preparation method uses chemical synthesis method: PGII antigen epitope peptides (1) and (2) are synthesized respectively by solid-phase method using the American ABI431A automatic peptide synthesizer. The purity of the epitope peptide was assessed by high performance liquid chromatography, and the concentration of the peptide was determined. The molecular weights of the antigenic epitope peptides (1) and (2) of the present invention are 1299.66 and 1943.41 respectively, which are determined by mass spectrometry, and the synthesized polypeptide sequences are identified by polypeptide sequence determination.

[0109] 1. Synthesis of PGII epitope peptides (1) and (2)

[0110]The above peptides were synthesized by solid-phase method. The main idea of ​​solid-phase peptide synthesis is: first connect the carboxyl group of the carboxyl-terminal amino acid of the peptide chain to be synthesized with a...

Embodiment 2

[0197] Embodiment 2: the PGII antigen epitope peptide (1) and (2) obtained in Example 1 are linked with carrier protein respectively to prepare PGII antigen (1) and (2), utilize gained antigen (1) and (2) respectively Animals are immunized to prepare specific monoclonal and polyclonal antibodies using the antigen (1), and specific monoclonal and polyclonal antibodies are prepared using the antigen (2).

[0198] 1. Antigen preparation: The PGII peptides (1) and (2) were respectively linked with the carrier protein KLH (keyhole limpet hemocyanin) by the BDB (Bis-diazotizedbenzidinedichloride) method to prepare PGII antigens (1) and (2).

[0199] Take PGII peptide (1) or (2) 10.0mg, dissolve with 1ml0.1MPBS buffer (pH7.4); KLH10mg, dissolve with 0.2M borate buffer (pH9.0) 20ml; then mix the two , cooled to 0°C, take BDBCl 2 110 μL, reacted at room temperature for 1.5 h, dialyzed overnight, then aliquoted, and stored at -20 °C.

[0200] In the present embodiment, the formula of ...

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Abstract

The invention relates to an immunochromatography test paper for detecting a human PGII protein and a preparation method thereof. The test paper employs a double antibody sandwich method for detection of the human PGII protein. The double antibody sandwich method uses a first PGII monoclonal antibody labeled with fluorescent microspheres as a capture antibody, and the first PGII monoclonal antibody is one of the sequences from the SEQ ID NO.1 and SEQ ID NO.2 in the sequence table; and the double antibody sandwich method uses a second PGII monoclonal antibody as a detection antibody, and the second PGII monoclonal antibody is another one from SEQ ID NO.1 and SEQ ID NO.2 in the sequence table. The invention of the immunochromatography test paper has the advantages of simple operation, rapidness, wide detection range, high specificity and good sensitivity.

Description

technical field [0001] The present invention belongs to the field of polypeptide chemistry and immunology, and specifically relates to human pepsinogen II (PGII) epitope peptide, PGII specific antigen prepared by using the epitope peptide and corresponding monoclonal antibody or polyclonal antibody, said Use of antibody in preparing human PGII in vitro diagnostic kit, human PGII in vitro diagnostic kit, fluorescent immunochromatographic test paper for quantitative detection of human PGII protein in analyte and preparation method thereof. Background technique [0002] Pepsinogen (PG) is the inactive precursor of pepsin in gastric juice. Human pepsinogen can be divided into seven isozymes from PG1 to PG7 according to its electrophoretic mobility from fast to slow, and can be divided into PGI and PGII according to its biochemical properties, immunogenicity, cell source and tissue distribution. a subgroup. The immunogenicity of PG1 to PG5 among the seven isoenzymes is similar,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/533
CPCG01N33/531G01N33/533G01N33/57446G01N33/68
Inventor 朱建安
Owner 朱建安
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