Hybridoma cell line C1 capable of secreting anti-paromomycin monoclonal antibody and application of hybridoma cell line C1

A hybridoma cell line and a paromomycin-resistant technology, applied in the field of immunochemistry, can solve the problem of no paromomycin ELISA method and the like

Active Publication Date: 2017-03-22
DELISI GROUP +1
View PDF1 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most aminoglycoside antibiotics have corresponding ELIS

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
  • Hybridoma cell line C1 capable of secreting anti-paromomycin monoclonal antibody and application of hybridoma cell line C1
  • Hybridoma cell line C1 capable of secreting anti-paromomycin monoclonal antibody and application of hybridoma cell line C1
  • Hybridoma cell line C1 capable of secreting anti-paromomycin monoclonal antibody and application of hybridoma cell line C1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Example 1: Synthesis of Immunogens and Coating Gens

[0014] The immunogen was coupled by the glutaraldehyde method: 31.96 mg of paromomycin was dissolved in 4 mL of 0.01 M phosphate buffer solution, 200 μL of 1% glutaraldehyde aqueous solution was added and stirred for 15 min. 20 mg of bovine serum albumin was dissolved in 2 mL of 0.01M phosphate buffer solution, reacted at 4°C for 1 h, added an appropriate amount of sodium borohydride to terminate the reaction, and incubated at 4°C for 2 h. The reaction mixture was dialyzed to remove free paromomycin to obtain the immunogen. The coating was synthesized by the carbodiimide method: 10 mg ovalbumin was dissolved in 1 mL pH 4.7 0.1 M MES (morpholine ethanesulfonic acid) buffer solution, 2 mg EDC and 1.2 mg NHS were added, and reacted at room temperature for 2 h. 19.03 mg paromomycin dissolved in 1 mL 0.05M KH 2 PO 4 , the activated protein solution was slowly added dropwise into the paromomycin solution, and reacted fo...

Embodiment 2

[0015] Example 2: Immunization of mice

[0016] The immunogen was diluted to an appropriate concentration with physiological saline, emulsified with an equal volume of Freund's adjuvant, and injected at multiple points on the back of 6-8 week-old BALB / c mice. For the first immunization, complete Freund's adjuvant was used at a dose of 100 μg. The following four booster immunizations used incomplete Freund's adjuvant, the immunization dose was 50 μg, and the immunization interval was 21 days. After the third immunization, blood was collected from the tail vein of the mice, and the sera were tested by indirect ELISA. After the fifth immunization, the mice with high titer and inhibition were screened for intraperitoneal immunization, and the spleens of the mice were taken out three days later for cell fusion.

Embodiment 3

[0017] Example 3: Cell Fusion and Screening

[0018] Mouse splenocytes and tumor cells were fused under the action of PEG 1500 to form hybridoma cells. Detection was carried out on the seventh day after fusion, and the cell lines with high titer and good paromomycin recognition were screened for subcloning, and so on, 5 times of subcloning were performed to obtain pure cell lines.

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 provides a hybridoma cell line C1 capable of secreting an anti-paromomycin monoclonal antibody and an application of the hybridoma cell line C1 and belongs to the technical field of immunochemistry. A mouse monoclonal cell line C1 is prepared with a conventional cell fusion technology, and is preserved in the CGMCC (China General Microbiological Culture Collection Center) with the preservation number being CGMCC No. 12025. The monoclonal antibody secreted by the cell line is determined by indirect competitive enzyme-linked immunosorbent assay, and the IC50 (half maximal inhibitory concentration) for paromomycin and the IC50 for neomycin are 0.61 mu g / L and 2.43 mu g / L respectively. The recovery rate range for paromomycin and neomycin in animal food is 64.56%, 105.85% and 54.08%-100.55%. Raw materials are provided for immunoassay of paromomycin residues in animal food, and the monoclonal antibody has practical application value.

Description

technical field [0001] The invention relates to a mouse-derived hybridoma cell strain C1 and the secreted monoclonal antibody capable of recognizing paromomycin, which can be used for the detection of paromomycin residues in food and belongs to the technical field of immunochemistry. Background technique [0002] Paromomycin is a broad-spectrum aminoglycoside antibiotic secreted by Streptomyces. The structure of paromomycin is very similar to neomycin, the only difference is that the amino group at position 6 of neomycin is replaced by a hydroxyl group. The antibacterial spectrum of paromomycin is similar to that of kanamycin, and it has a strong killing effect on amoeba, and it also has antibacterial activity against some protozoa, including: Leishmania, amoeba dysenteriae, cryptosporidium insect. Clinically, paromomycin is used to treat amoebiasis, giardiasis and leishmaniasis. In animal husbandry, it is used to treat bacterial infections, including histomoniasis of t...

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
IPC IPC(8): C12N5/20C07K16/44G01N33/577G01N33/543
CPCC07K16/44
Inventor 徐丽广陈燕妮胥传来匡华刘丽强宋珊珊吴晓玲郑乾坤
Owner DELISI GROUP
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