Immunochromatography test paper for detecting human PGI protein and preparation method thereof

A technology of fluorescent immunochromatography and test paper, which is applied in the field of peptide chemistry and immunology, and achieves the effects of high specificity, wide detection range and good sensitivity

Active Publication Date: 2015-03-18
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 PGI protein

Method used

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  • Immunochromatography test paper for detecting human PGI protein and preparation method thereof
  • Immunochromatography test paper for detecting human PGI protein and preparation method thereof
  • Immunochromatography test paper for detecting human PGI protein and preparation method thereof

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preparation example Construction

[0055] The preparation method of the PGI epitope peptide of the present invention can be a chemical synthesis method: the antigen 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 1657.24 and 1616.93 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.

[0056] 2. PGI antigen

[0057] The present invention also provides a PGI antigen prepared by coupling one of the human PGI epitope peptides (1) and (2) of the present invention with a carrier protein. Specifically, the present invention provides PGI antigens (1) and (2), whe...

Embodiment 1

[0108] Example 1: Preparation of PGI epitope peptides (1) and (2).

[0109] The preparation method uses chemical synthesis method: the PGI 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 1657.24 and 1616.93 respectively, which are determined by mass spectrometry, and the synthesized polypeptide sequences are identified by polypeptide sequence determination.

[0110] 1. Synthesis of PGI epitope peptides (1) and (2)

[0111] 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...

Embodiment 2

[0198] Example 2: The PGI antigen epitope peptides (1) and (2) obtained in Example 1 are linked to carrier protein to prepare PGI antigens (1) and (2), respectively, and the obtained antigens (1) and (2) are used to 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).

[0199]1. Antigen preparation: PGI peptides (1) and (2) were respectively linked with carrier protein KLH (keyhole limpet hemocyanin) by BDB (Bis-diazotizedbenzidine dichloride) method to prepare PGI antigens (1) and (2) .

[0200] Take 10.0mg of PGI peptide (1) or (2), dissolve it with 1ml 0.1M PBS buffer (pH7.4); dissolve KLH10mg with 20ml of 0.2M borate buffer (pH9.0); then mix the two Mix, cool 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.

[0201] In the present embodiment, the formula of PBS ...

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Abstract

The invention relates to a Immunochromatography test paper for detecting a human PGI protein and a preparation method thereof. The test paper employs a double antibody sandwich method for detection of the human PGI protein. The double antibody sandwich method uses a first PGI monoclonal antibody labeled with fluorescent microspheres as a capture antibody, and the first PGI 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 PGI monoclonal antibody as a detection antibody, and the second PGI 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 I (PGI) epitope peptide, PGI-specific antigen prepared by using the epitope peptide and corresponding monoclonal antibody or polyclonal antibody, said The application of the antibody in the preparation of a human PGI in vitro diagnostic kit, the human PGI in vitro diagnostic kit, and a fluorescent immunochromatographic test paper for quantitatively detecting human PGI protein in analytes and a 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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N21/64
CPCG01N33/57446G01N33/68
Inventor 朱建安
Owner 朱建安
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