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Colloidal gold immunochromatographic test paper capable of quickly detecting soybean Kunitz trypsin inhibiting factor and preparation method

A technology of immunochromatographic test paper and colloidal gold, which is applied in the direction of biological testing, anti-plant immunoglobulin, immunoglobulin from serum, etc., to achieve the effect of displaying image, judging image, not easy to false positive and false negative

Active Publication Date: 2015-09-16
HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But there is still no report on the detection method of KTI immunochromatographic test paper, so this method will become a development direction of the detection method of soybean trypsin inhibitor

Method used

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  • Colloidal gold immunochromatographic test paper capable of quickly detecting soybean Kunitz trypsin inhibiting factor and preparation method
  • Colloidal gold immunochromatographic test paper capable of quickly detecting soybean Kunitz trypsin inhibiting factor and preparation method

Examples

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preparation example Construction

[0022] The preparation process of the test paper of the present invention comprises: the preparation of KTI rabbit polyclonal antibody, the preparation of KTI mouse monoclonal antibody, the preparation of binding pad, the preparation of sample pad, the preparation of cellulose film and the assembly of test paper and the like.

[0023] 1. Preparation of KTI rabbit-derived polyclonal antibody:

[0024] Clean grade 3-month-old New Zealand white rabbits were immunized with KTI solution and injected subcutaneously in the neck at 4-6 points. The first immunization dose was 200 μg / monkey, diluted KTI with sterile PBS and mixed with an equal amount of Freund’s complete adjuvant (FCA), and fully emulsified at 25000 r / min for 10 minutes; the booster immunization dose was increased to 300 μg / bird, and immunized 4 times with Dilute KTI in sterile PBS and mix with an equal amount of Freund's incomplete adjuvant (FIA), fully emulsify at 25000r / min for 10min. A total of 4 times of immunizat...

Embodiment 1

[0043] Embodiment one: see figure 1 and figure 2 . Among the figure 1 is the support layer, made of plastic sheet strips; 2 is the sample pad, made of glass fiber cotton; 3 is the binding pad, the glass fiber cotton of the KTI mouse monoclonal antibody labeled with colloidal gold nanoparticles is adsorbed; 4 is a cellulose membrane, which adopts a nitrocellulose membrane; the absorbent pad 5 at the handle end is made of absorbent filter paper. The sample pad 2, the bonding pad 3, the cellulose membrane 4, and the water-absorbent pad 5 are pasted and fixed on the support layer 1 in sequence from left to right, and the fibers at the junctions overlap each other. On the cellulose membrane 4, there is an invisible detection line 6, which is made of KTI rabbit-derived polyclonal antibody solution; the invisible quality control line 7 is made by spraying a goat anti-mouse IgG antibody solution on the cellulose membrane, two lines The bands are arranged in parallel to form a comb...

Embodiment 2

[0049] Embodiment 2: The structure of the test paper is basically the same as that of Embodiment 1, except that the sample pad 2 is made of nylon film, and the cellulose film 4 is made of pure cellulose film.

[0050] Feed testing: first crush the sample, pass through a 60-mesh sieve, and then degrease the crushed sample with petroleum ether. The subsequent sample preparation method is the same as in Example 1.

[0051] The operation method and result judgment are the same as in the first embodiment.

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Abstract

The invention relates to a colloidal gold immunochromatographic test paper capable of quickly detecting a soybean Kunitz trypsin inhibiting factor and a preparation method. The test paper detects a KTI (Key Technology Index) based on a double-antibody sandwich principle, the test paper container comprises a support layer, an adsorption layer and a protection layer, and the adsorption layer is sequentially provided with a sample cushion, a binding cushion, a cellulose membrane and a water adsorption cushion from a test end. The binding cushion absorbs a specific antibody with the KTI, the cellulose membrane is provided with a detection line used for a KTI anti-cleaved caspase rabbit polyclonal antibody solution spray point and a quality control line used for an anti-mouse IgG (immunoglobulin G) antibody or a rabbit anti-mouse IgG antibody solution spray point, and the KTI specific antibody adopts a KTI murine monoclonal antibody marked by colloidal gold nano-particles. The immunochromatographic test paper provided by the invention is strong in specificity, high in flexibility, convenient, intuitionistic and accurate to detect, low in cost, wide in scope of application, and easy for popularization and application.

Description

technical field [0001] The invention relates to a colloidal gold immunochromatographic test paper for rapidly detecting soybean Kunitz trypsin inhibitory factor and a preparation method thereof. Background technique [0002] Soybean is rich in protein, generally 35%~38%, with balanced amino acid composition and rich in unsaturated fatty acids. It is an important source of vegetable protein and vegetable oil for human and animal bodies. Due to its wide planting area and large output, it has been widely used as raw material for food and feed processing for a long time, and has extremely high use value for human health and animal nutrition. However, soybean also contains a variety of anti-nutritional factors, which largely affect the development and utilization of soybean. [0003] Soybean trypsin inhibitor (Kunitz trypsin inhibitor, KTI) is one of the main anti-nutritional factors of soybean. 6%. KTI was first isolated from soybean by Kunitz in 1945, its molecular weight is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/558G01N33/577G01N33/532C07K16/16C07K16/06
CPCC07K16/06C07K16/16G01N33/532G01N33/558G01N33/577G01N33/68G01N33/6803G01N2333/81
Inventor 邓瑞广胡骁飞王耀李青梅王方雨杨继飞郝天红
Owner HENAN ACAD OF AGRI SCI
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