Fluorescent immunochromatographic test paper and preparation method for joint detection of human pgi protein and human pgii protein

A fluorescent immunochromatography, protein technology, applied in biological testing, fluorescence/phosphorescence, measurement devices, etc., can solve the problem of not combined detection of human PGI protein fluorescent immunochromatography test strips, etc., to achieve good antigenicity, wide detection range, Easy to use effect

Active Publication Date: 2016-05-18
朱建安
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no fluorescent immunochromatographic test strip for the combined detection of human PGI protein and human PGII protein

Method used

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  • Fluorescent immunochromatographic test paper and preparation method for joint detection of human pgi protein and human pgii protein
  • Fluorescent immunochromatographic test paper and preparation method for joint detection of human pgi protein and human pgii protein
  • Fluorescent immunochromatographic test paper and preparation method for joint detection of human pgi protein and human pgii protein

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preparation example Construction

[0066] 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.

[0067] 2. PGI antigen

[0068] 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

[0148] Example 1: Preparation of PGI epitope peptides (1) and (2) and PGII epitope peptides (3) and (4).

[0149] The preparation method uses chemical synthesis method: using the American ABI431A automatic peptide synthesizer, the PGI antigenic epitope peptides (1) and (2) and the PGII antigenic epitope peptides (3) and (4) are synthesized respectively by a solid-phase method. 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 PGI antigenic epitope peptides (1) and (2) of the present invention are 1657.24 and 1616.93 respectively, and the molecular weights of the PGII antigenic epitope peptides (3) and (4) are respectively 1299.66 and 1943.41, which are determined by mass spectrometry. Sequencing identifies the synthesized polypeptide sequence.

[0150] 1. Synthesis of PGI epitope peptides (1) and (2) and PGII epitope peptides (3) and (4)

[0151] The abov...

Embodiment 2

[0241] Embodiment 2: The PGI epitope peptide (1) and (2) and PGII antigen epitope peptide (3) and (4) obtained in Example 1 are connected with carrier protein respectively to prepare PGI antigen (1), PGI antigen (2), PGII antigen (3) and PGII antigen (4), utilize the obtained antigen (1), (2), (3) and (4) to immunize animals respectively, thereby utilize antigen (1), (2) respectively , (3) and (4) prepare specific monoclonal antibodies and polyclonal antibodies.

[0242] 1. Antigen preparation: PGI peptide (1), PGI peptide (2), PGII peptide (3) and PGII peptide (4) were combined with carrier protein KLH (keyhole pore) by BDB (Bis-diazotizedbenzidinedichloride) method Hemocyanin) was connected to prepare PGI antigen (1), PGI antigen (2), PGII antigen (3) and PGII antigen (4).

[0243] Take 10.0mg of PGI peptide (1) or PGI peptide (2) or PGII peptide (3) or PGII peptide (4), dissolve it in 1ml 0.1MPBS buffer (pH7.4); KLH10mg, use 0.2M boron Dissolve 20ml of salt buffer solutio...

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Abstract

The present invention relates to fluorescent immunochromatography test paper for combined detection of human PGI protein and human PGII protein, and a production method. According to the test paper of the present invention, human PGI protein and human PGII protein are detected through a double-antibody sandwich method, wherein the capture antibodies adopted by the double-antibody sandwich method respectively are first PGI monoclonal antibody derived from one of the sequence 1 and the sequence 2 in the sequence table and first PGII monoclonal antibody derived from one of the sequence 3 and the sequence 4 in the sequence table, and the detection antibodies adopted by the double-antibody sandwich method respectively are second PGI monoclonal antibody derived from the other one of the sequence 1 and the sequence 2 in the sequence table and second PGII monoclonal antibody derived from the other one of the sequence 3 and the sequence 4 in the sequence table; and the fluorescent immunochromatography test paper of the present invention ha characteristics of convenient and rapid operation, 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 Use of antibody in preparation of human PGI in vitro diagnostic kit, human PGI in vitro diagnostic kit, and human pepsinogen II (PGII) epitope peptide, PGII specific antigen prepared with the antigenic epitope peptide and corresponding single Cloned antibody or polyclonal antibody, the purposes of said antibody in the preparation of human PGII in vitro diagnostic kit, human PGII in vitro diagnostic kit, the present invention also relates to the method for combined quantitative detection of human PGI protein and human PGII protein in analyte A fluorescent immunochromatographic test paper and a preparation method thereof. Background technique [0002] Pepsinogen (PG) is the inac...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N21/64G01N33/531G01N33/533
CPCG01N33/531G01N33/533G01N33/57446G01N33/68
Inventor 朱建安
Owner 朱建安
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