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Preparation and application of vaccine immunity effect evaluation protein chip

A technology of protein chips and vaccines, which is applied in the field of preparation of protein chips for vaccine immune effect evaluation, can solve the problems of time-consuming detection methods, large-scale and effective evaluation of vaccine immune effect antibody levels, and reduced evaluation efficiency, so as to promote research and development Application, high-throughput detection, and efficient evaluation of tool effects

Inactive Publication Date: 2011-09-07
INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented device allows researchers to test how well an organism's defense against disease caused by bacteria infectious agents like viruses. By attaching tiny particles called chirp onto small carriers made from glass fiber clothings (fiberglass), they were able to attach these minute particles directly into their blood vessels without affecting them physically. These devices have been developed over several decades ago but still lacked any technical features related to testing protection levels between two types of virus strains - one containing both protections while another harmlessly free from such measures.

Problems solved by technology

Technologically speaking, current testing tools involve costly procedures like culture assays and flow-cytometric measurements, leading to slow down progress towards more accurate identification of relevant strains of viruses involved in future epidemic situations. There is therefore a demand for faster and higher-quality diagnoses without these drawbacks associated with conventional approaches.

Method used

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  • Preparation and application of vaccine immunity effect evaluation protein chip
  • Preparation and application of vaccine immunity effect evaluation protein chip
  • Preparation and application of vaccine immunity effect evaluation protein chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Treatment of Partial Disease Pathogen Protective Antigens

[0036] 1. Materials

[0037] Group A meningococcal polysaccharide vaccine was purchased from Beijing Tiantan Biological Products Co., Ltd.; Bacillus anthracis lysate was donated by Professor Wang Xiliang; live attenuated polio vaccine sugar pills were purchased from the Institute of Medical Biology, Chinese Academy of Medical Sciences; Live viruses (Types I, II, III) were obtained from the 302 Hospital of the People’s Liberation Army; Adsorbed Diphtheria-Tetanus Vaccine was purchased from Shanghai Institute of Biological Products; MMR combined live attenuated vaccine was purchased from Beijing Tiantan Biological Products Co., Ltd.; The glass sheet base was purchased from CEL Associates; Cy3-labeled goat anti-human IgG was purchased from sigma.

[0038] 2. Methods and Results

[0039] 1. Comparison of the effects of different methods for treating group A meningococcal polysaccharide vaccine

[0040]...

Embodiment 2

[0059] Example 2 National immunization program disease protective antibody detection protein chip probe lattice design and chip preparation

[0060] 1. Materials

[0061] HBsAg was expressed and preserved in our laboratory; measles virus H protein, rubella virus E1 protein, mumps virus NP protein, hepatitis A virus vp1 and vp3 proteins, Japanese encephalitis virus E protein, Mycobacterium tuberculosis Ag85B antigen were purchased from abcam company; IgG, BSA, pertussis Bacillus FHA, diphtheria toxin, and tetanus toxin were purchased from Sigma; live attenuated polio vaccine sugar pills were purchased from the Institute of Medical Biology, Chinese Academy of Medical Sciences; group A meningococcal polysaccharide vaccine was purchased from Beijing Tiantan Biological Products Co., Ltd. The NP protein of hemorrhagic fever virus was donated by Professor Liang Mifang; the Bacillus anthracis PA protein was purified by our laboratory; the Leptospira antigen was donated by Professor Du...

Embodiment 3

[0072] Example 3 Protein Chip Detection of Protective Antibodies in Serum Samples

[0073] 1. Materials

[0074] Cy3-labeled goat anti-human IgG, nano-gold-labeled goat anti-human IgG, and HRP-labeled goat anti-human IgG were purchased from Sigma; nano-gold composite substrate (Nanogold-DAB), silver enhancement reagents A and B were prepared and stored in our laboratory.

[0075] 2. Methods and Results

[0076] Take the protein chip prepared in the above example and equilibrate to room temperature, dilute the serum sample 1:5 with sample diluent 0.01mol / L PBS (25% FCS, pH=7.4), add 10 microliters to each reaction zone, The chip was incubated at 37°C for 30 minutes. Wash the chip with washing solution PBST (0.01 mol / L, 0.05% Tween-20, pH=7.4) gently for 3 times, 20 seconds each time, and prepare for color development after drying.

[0077] 1. Fluorescent color development

[0078] Cy3-labeled goat anti-human IgG was diluted 1:400, 10 microliters were added to each reaction ...

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Abstract

The invention relates to a preparation and application of a vaccine immunity effect evaluation protein chip. By using the invention, whether corresponding protective antibodies are contained in bodies of vaccinated people or animals or not or whether the vaccines have protective effects after the people or animals are vaccinated for a period of time or not can be detected. In the invention, the protein chip is prepared on a modified substrate by using protective antigens of multiple disease pathogens or antigen material points of corresponding vaccines after being treated by different methods, protective antibodies of the multiple vaccines can be simultaneously detected through an experiment, and multiple specimens can be simultaneously analyzed; and multiple different signal detecting methods are obtained; the vaccine immunity effect evaluation protein chip provided by the invention has the advantages of speediness, accuracy, high throughput, high sensitivity and high specificity; and a new technology and a new product can be provided for the immunity effect evaluation of the vaccine.

Description

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Claims

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Application Information

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Owner INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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