Double-antibody sandwich enzymelinked immunosorbent detection kit and preparation method thereof

An enzyme-linked immunoassay detection and kit technology, which is applied in the field of kits to achieve the effects of high specificity, cost reduction, and high detection sensitivity

Active Publication Date: 2012-05-02
GUANGDONG PHARMA UNIV
View PDF4 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003]The joint detection of ELISA kits currently used is to operate the enzyme plates of several markers at the same time, which has achieved the purpose of joint detection. This operation will large human error

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Double-antibody sandwich enzymelinked immunosorbent detection kit and preparation method thereof
  • Double-antibody sandwich enzymelinked immunosorbent detection kit and preparation method thereof
  • Double-antibody sandwich enzymelinked immunosorbent detection kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Detection of NSE and CK19 Antigens by Double Antibody Sandwich ELISA Kit

[0022] (1) Add standard products of various concentrations to the reaction wells, add 0 ng / mL, 7.5 ng / mL, 31.15 ng / mL, 125 ng / mL, 250 ng / mL, 500 ng / mL and the serum to be tested in each well, and wait for The tested serum was diluted 5 times, 100uL was added to each well, and placed at 37°C for 1h.

[0023] (2) Pat dry the liquid in the plate, add washing solution to wash the plate, 200uL per well, 30s each time, wash 5 times.

[0024] (3) Diluted enzyme-labeled antibody was added to each well, 100uL per well, incubated at 37°C for 1h.

[0025] (4) Pat dry the liquid in the plate, add washing solution to wash the plate, 200uL per well, 30s each time, wash 5 times.

[0026] (5) Add chromogenic solution PNPP to each well, 100uL per well, react at 37°C for 10 min, add stop solution, and read at 405nm.

[0027] (6) Pat dry the liquid in the plate, add washing solution to wash the plate...

Embodiment 2

[0029] Example 2 Repeatability analysis of double-antibody sandwich ELISA method

[0030] Positive serum samples were selected and tested with the kit obtained in Example 1, and each sample was tested in parallel for 3 wells, and the intra-batch repeatability of the kit was tested.

[0031] Table 1 Alkaline phosphatase (AP) labeled NSE

[0032]

[0033] Positive serum samples were selected and tested with 3 different batches of the double-antibody sandwich diagnostic kit prepared in Example 1, and the inter-batch reproducibility of the kit was tested.

[0034]

[0035]

Embodiment 3

[0036] Embodiment 3 kit stability analysis

[0037] Store the kit in an environment of 37°C, and test it regularly until the maximum value of its quality index drops by half. It is generally considered that storing at 37°C for one day is equivalent to storing at 4-10°C for 1.5 months. For example, if the analysis kit is placed at 37°C for 6 days, the OD value detected by NSE will drop by half on the 6th day, which means it can be stored at 4°C for more than 9 months. The detected OD value of CK19 will drop by half on the 4th day. Because CK19 can be stored at 4°C for more than 6 months. Therefore, the kit can be stored at 4°C for at least 6 months.

[0038]

[0039]

[0040]

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a double-antibody sandwich enzymelinked immunosorbent detection kit and a preparation method of the double-antibody sandwich enzymelinked immunosorbent detection kit. The detection kit comprises a perforated plate coated with neuron-specific enolase (NSE) monoclonal antibodies and CK 19 (creatine kinase 19) monoclonal antibodies, NSE labeled by horse radish peroxidase (HRP), CK 19 labeled by alkaline phosphatase, chromogenic substrate tetramethylbenzidine (TMB), p-nitrophenyl disodium phosphate (PNPP), disodium hydrogen phosphate and substrate stop solution. The double-antibody sandwich enzymelinked immunosorbent detection kit has the advantages of being simple and convenient, sensitive, stable, good in repeatability and the like, and has practical value.

Description

technical field [0001] The invention relates to the field of kits, in particular to a double-antibody sandwich enzyme-linked immunoassay kit and a preparation method thereof. Background technique [0002] Lung cancer is one of the most common malignant tumors with the largest number of deaths in the world, and its incidence rate is also the highest among all malignant tumors, because the onset of lung cancer is hidden. At present, there is still a lack of effective screening and early diagnosis methods. Patients with symptoms are mostly in the late stage, and the prognosis is poor. The total five-year survival rate is less than 15%, and the symptoms are less than 10%. However, in early-diagnosed lung cancer patients, surgery The prognosis of the treatment is called that the advanced lung cancer has been significantly improved, and its survival rate can reach 70%. Traditional clinical diagnostic methods such as chest X-ray, bronchoscopy, sputum cytology and other methods. ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
Inventor 臧林泉刘大路潘雪刁
Owner GUANGDONG PHARMA UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap