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Method for identifying silk fabric residues based on enzyme-linked immunosorbent assay

An enzyme-linked immunization and silk fabric technology, applied in the field of cultural relic detection, can solve the problems of increased interference, increased sample impurities, and reduced content of silk fabric fragments, and achieves the effects of low production cost, simple process and small molecular weight.

Active Publication Date: 2019-03-29
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The fabric content of ancient silk fabric fragments decreases accordingly, and the sample impurities increase and interference increases. If we continue to use traditional methods, it will be difficult to make accurate judgments. How to find a more scientific method to identify ancient silk fabrics has always been a cultural relic. conundrum in conservation

Method used

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  • Method for identifying silk fabric residues based on enzyme-linked immunosorbent assay

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Experimental program
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Effect test

Embodiment 1

[0040] 1) Preparation of silk-specific antibody: Weigh mulberry silk and tussah silk, degumming, dissolving, dialysis, purification and freeze-drying to obtain silk fibroin powder. The silk fibroin powder was dissolved in PBS and injected into SPF grade Balb / C purebred female mice. After several spleen immunizations, the spleen cells were fused with myeloma cells. After cloning, screening, culturing and purification, respectively, The monoclonal antibody to mulberry silk fibroin (McAb-BM) and the monoclonal antibody to tussah silk fibroin (McAb-AP) were obtained.

[0041] 2) Preparation of antibody Fab fragments: Select immobilized papain to prepare Fab fragments by enzymatic hydrolysis. Activate the immobilized papain in the separation column, add 0.25 mL papain solution to the 0.8 mL separation column, put the separation column into a 2 mL centrifuge tube, centrifuge at 10000 rpm for 1 min, discard the centrifuged protein and store solution, add 0.5 mL 20mM cysteine ​​diges...

Embodiment 2

[0051] 1) Preparation of silk-specific antibody: Weigh mulberry silk and tussah silk, degumming, dissolving, dialysis, purification and freeze-drying to obtain silk fibroin powder. The silk fibroin powder was dissolved in PBS and injected into SPF grade Balb / C purebred female mice. After several spleen immunizations, the spleen cells were fused with myeloma cells. After cloning, screening, culturing and purification, the obtained Mulberry silk fibroin monoclonal antibody (McAb-BM) and tussah silk fibroin monoclonal antibody (McAb-AP).

[0052]2) Preparation of antibody Fab fragments: Select immobilized papain to prepare Fab fragments by enzymatic hydrolysis. Activate the immobilized papain in the separation column, add 0.25 mL papain solution to the 0.8 mL separation column, put the separation column into a 2 mL centrifuge tube, centrifuge at 10000 rpm for 1 min, discard the centrifuged protein and store solution, add 0.5 mL 20mM cysteine ​​digestion solution, repeat centrifu...

Embodiment 3

[0061] 1) Preparation of silk-specific antibody: Weigh mulberry silk and tussah silk, degumming, dissolving, dialysis, purification and freeze-drying to obtain silk fibroin powder. The silk fibroin powder was dissolved in PBS and injected into SPF grade Balb / C purebred female mice. After several spleen immunizations, the spleen cells were fused with myeloma cells. After cloning, screening, culturing and purification, the obtained Mulberry silk fibroin monoclonal antibody (McAb-BM) and tussah silk fibroin monoclonal antibody (McAb-AP).

[0062] 2) Preparation of antibody Fab fragments: Select immobilized papain to prepare Fab fragments by enzymatic hydrolysis. Activate the immobilized papain in the separation column, add 0.25 mL papain solution to the 0.8 mL separation column, put the separation column into a 2 mL centrifuge tube, centrifuge at 10000 rpm for 1 min, discard the centrifuged protein and store solution, add 0.5 mL 20mM cysteine ​​digestion solution, repeat centrif...

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Abstract

The invention relates to the technical field of cultural relic detection, and discloses a method for identifying silk fabric residues based on enzyme-linked immunosorbent assay. The method comprises the following steps: preparing monoclonal antibodies of a mulberry silk fibroin protein and a tussah silk fibroin protein; treating the antibodies using an immobilized papain to obtain Fab fragment antibodies with higher activity; and compounding the fragment to a gold nanoparticle to obtain a specific Fab-Au NP composite. The enzyme-linked immunosorbent assay shows that the composite can efficiently identify the varieties of ancient silk fabric residues. The method is mild in reaction, environmentally friendly and harmless; and when detecting ancient silk fabrics, the method has the characteristics of small sample consumption, intuitional, fast and high sensitivity.

Description

technical field [0001] The invention relates to the technical field of cultural relic detection, in particular to a method for identifying silk fragments based on ELISA. Background technique [0002] As an important symbol and carrier of Chinese civilization, silk carries rich cultural, technological and social connotations, leading the ancient Chinese civilization from the Silk Road to the world, and has made an indelible contribution to the development of human civilization in the world. [0003] In 1958, a batch of silk threads, ribbons and uncarbonized silk pieces were unearthed from the Yangliangzhu cultural site in Qianshan, Zhejiang; in the 1980s, fragments of silk and linen fabrics were found at the Neolithic site in Qingtai Village, Xingyang, Henan. Many scholars at home and abroad have analyzed the silk residues unearthed from archaeological excavations in order to discover earlier silk. In the excavation of ancient silk, due to the influence of factors such as ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/535G01N33/543
CPCG01N33/535G01N33/54346G01N33/577
Inventor 陈茹茹王秉胡铭周朱诚胡智文
Owner ZHEJIANG SCI-TECH UNIV
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