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Stable and sensitive type IV collagen (IV-C) detection reagent

A collagen and detection reagent technology, applied in the field of type IV collagen (IV-C) detection, can solve the problems of difficult to popularize and use, radiation exposure, time-consuming and labor-intensive, etc., to meet clinical needs, strong anti-interference, and easy to use Effect

Active Publication Date: 2018-09-04
BIOBASE BIODUSTRY (SHANDONG) CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the enzyme-linked immunoassay (ELISA) requires specific equipment, is time-consuming and laborious, and is not easy to promote and use; while radioimmunoassay technology has high sensitivity, but the equipment is expensive, and there are problems such as radiation and pollution.

Method used

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  • Stable and sensitive type IV collagen (IV-C) detection reagent
  • Stable and sensitive type IV collagen (IV-C) detection reagent
  • Stable and sensitive type IV collagen (IV-C) detection reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] The type IV collagen detection reagent described in this embodiment includes reagent R1 and reagent R2:

[0048] 1) The components of reagent R1 are:

[0049] TAPSO buffer (pH=7.2, 25°C) 80mmol / L,

[0050] RHEODOL TW-O320V (polyoxyethylene sorbitan trioleate) 1g / L,

[0051] AMINON C-11S (Methyl Cocoamide) 0.5g / L,

[0052] preservative 0.1g / L;

[0053] 2) The components of reagent R2 are:

[0054] TAPSO buffer (pH=7.5, 25°C) 50mmol / L,

[0055] Goat anti-human collagen IV antibody coated latex particles 0.12%,

[0056] polyvinylpyrrolidone 0.1g / L,

[0057] γ-cyclodextrin 0.2g / L,

[0058] xanthan gum 0.1g / L,

[0059] Sodium chloride 2g / L,

[0060] Glycerol 3ml / L,

[0061] Trehalose 15g / L,

[0062] PEG-6000 5g / L,

[0063] RHEODOL TW-O320V (polyoxyethylene sorbitan trioleate) 1g / L,

[0064] AMINON C-11S (Methyl Cocoamide) 0.5g / L,

[0065] preservative 0.1g / L.

[0066] 3) The method of using the reagents i...

Embodiment 2

[0069] The type IV collagen detection reagent described in this example includes reagent R1 and reagent R2 (the detection method is the same as that in Example 1):

[0070] 1) The components of reagent R1 are:

[0071] TAPSO buffer (pH=7.2, 25°C) 80mmol / L,

[0072] RHEODOL TW-O320V (polyoxyethylene sorbitan trioleate) 1g / L,

[0073] AMINON C-11S (Methyl Cocoamide) 0.5g / L,

[0074] preservative 0.1g / L;

[0075] 2) The components of reagent R2 are:

[0076] TAPSO buffer (pH=7.5, 25°C) 50mmol / L,

[0077] Goat anti-human collagen IV antibody coated latex particles 0.18%,

[0078] polyvinylpyrrolidone 0.1g / L,

[0079] γ-cyclodextrin 0.2g / L,

[0080] xanthan gum 0.1g / L,

[0081] Sodium chloride 2g / L,

[0082] Glycerol 3ml / L,

[0083] Trehalose 20g / L,

[0084] PEG-6000 8g / L,

[0085] RHEODOL TW-O320V (polyoxyethylene sorbitan trioleate) 1g / L,

[0086] AMINON C-11S (Methyl Cocoamide) 0.5g / L,...

Embodiment 3

[0089] The type IV collagen detection reagent described in this example includes reagent R1 and reagent R2 (the detection method is the same as that in Example 1):

[0090] 1) The components of reagent R1 are:

[0091] TAPSO buffer (pH=7.2, 25°C) 80mmol / L,

[0092] RHEODOL TW-O320V (polyoxyethylene sorbitan trioleate) 1g / L,

[0093] AMINON C-11S (Methyl Cocoamide) 0.5g / L,

[0094] preservative 0.1g / L;

[0095] 2) The components of reagent R2 are:

[0096] TAPSO buffer (pH=7.5, 25°C) 50mmol / L,

[0097] Goat anti-human collagen IV antibody coated latex particles 0.2%,

[0098] polyvinylpyrrolidone 0.1g / L,

[0099] γ-cyclodextrin 0.2g / L,

[0100] xanthan gum 0.1g / L,

[0101] Sodium chloride 2g / L,

[0102] Glycerol 3ml / L,

[0103] Trehalose 25g / L,

[0104] PEG-6000 10g / L,

[0105] RHEODOL TW-O320V (polyoxyethylene sorbitan trioleate) 1g / L,

[0106] AMINON C-11S (Methyl Cocoamide) 0.5g / L,

[0107] preservat...

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Abstract

The invention discloses a stable and sensitive type IV collagen (IV-C) detection reagent. The stable and sensitive type IV collagen (IV-C) detection reagent comprises a reagent R1 and a reagent R2; areaction system is optimized, TAPSO (N-[Tris(hydroxymethyl)metyl]-3-amino-2-hydroxypropanesulfonic acid) buffer is adopted, a plurality of stabilizing agents including polyvinylpyrrolidone, gamma-cyclodextrin, xanthan gum, NaCl, trehalose, PEG 6000, and glycerol are added, so that reagent stability is improved obviously; goat anti-human type IV collagen antibody coated latex particles are adopted,in detection, mutual crosslinking of adjacent latex particles is realized through antigen-antibody specific immunization reaction in detection, so that reagent sensitivity and accuracy are increasedobviously; and in addition, combination of novel nonionic surfactants RHEODOL, TW-O320V (tween 85) and AMINON C-11S (methyl cocoate alcohol amide) is adopted, so that antibody stability is promoted and maintained, system turbidity is avoided, and reagent stability is improved further.

Description

technical field [0001] The invention relates to the technical field of type IV collagen (IV-C) detection, in particular to a type IV collagen (IV-C) detection reagent and a detection method using the detection reagent. Background technique [0002] Type IV collagen (IV-C) is an important component of the basement membrane. The normal intrahepatic basement membrane mainly exists around blood vessels, lymphatic vessels, and bile ducts, and is absent in the sinusoidal wall of the liver. When liver disease occurs, inflammation develops, fibrous tissue hyperplasia is active, and a large amount of collagen deposits are generated. Various collagens increase, but the most important one is the increase of type IV collagen that constitutes the basement membrane. At present, it is believed that the determination of the concentration of serum type Ⅳ collagen can be used as a modern indicator for checking liver fibrosis. [0003] Serum IV-C concentration is related to the degree of liv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N35/00
CPCG01N33/6803G01N35/00
Inventor 李志明胡晓飞王美丽段洪东
Owner BIOBASE BIODUSTRY (SHANDONG) CO LTD
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