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Nano antibody for broad-spectrum recognition of Salmonella, recombinant vector, host cell and application of nano antibody

A nano-antibody and salmonella technology, applied in the field of molecular biology, can solve the problem of no recognition ability of salmonella, and achieve the effects of good detection results, high expression yield and good binding ability

Inactive Publication Date: 2021-04-27
NORTHWEST A & F UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the current prior art, the patent CN108864281B, which the applicant applied for and finally authorized on July 13, 2018, discloses a nanobody against Salmonella enteritidis and its application. However, the nanobody is mainly used in Salmonella enteritidis. For Other species of Salmonella have no ability to identify

Method used

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  • Nano antibody for broad-spectrum recognition of Salmonella, recombinant vector, host cell and application of nano antibody
  • Nano antibody for broad-spectrum recognition of Salmonella, recombinant vector, host cell and application of nano antibody
  • Nano antibody for broad-spectrum recognition of Salmonella, recombinant vector, host cell and application of nano antibody

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Experimental program
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Effect test

Embodiment

[0028] 1. Construction of anti-Salmonella phage display nanobody library

[0029] 1.1 Immunity of camels:

[0030] Select a healthy camel that has not been immunized with any antigen, and immunize the Bactrian camel with an equal volume of inactivated Salmonella enteritidis and the emulsified mixture of Freund's complete adjuvant at a concentration of 106cfu / mL. Freund's incomplete adjuvant was used to emulsify the antigen, and the immunization was boosted once every 2 weeks, for a total of 5 immunizations. One week after each immunization, the camel blood was collected to detect the antibody titer in the serum.

[0031] 1.2 Extraction of total RNA from blood: After the fifth immunization, the peripheral blood of camels was collected, and total RNA was extracted according to the operation steps of the RNA extraction kit.

[0032] 1.3 Reverse transcription to obtain cDNA:

[0033] The obtained total RNA is used as a template, oligo(dT) 15 The primers are used for reverse tr...

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Abstract

The invention discloses a nano antibody for broad-spectrum anti-salmonella of Salmonella. The amino acid sequence of the nano antibody is shown as SEQ ID NO: 1. The invention further provides a vector and host cell, containing the amino acid sequence of the nano antibody. The nano antibody can be well combined with salmonella enteritidis, salmonella typhimurium, salmonella hardahl, salmonella enteritidis and salmonella paratyphi B. At present, a monoclonal antibody and a polyclonal antibody are poor in stability, relatively high in preparation cost and easy to combine with surface protein of staphylococcus aureus, and the nano antibody for resisting salmonella provided by the invention is good in broad spectrum, strong in stability, high in expression yield and free of cross reaction with staphylococcus aureus. A double-nano antibody sandwich enzyme-linked immunoassay method established by the invention can be used for detecting various salmonella, and the detection result is relatively good. According to the invention, the problems of poor specificity and high cost of the existing detection method can be solved, so that the double-nano antibody sandwich enzyme-linked immunoassay method is more widely applied.

Description

technical field [0001] The invention belongs to the field of molecular biology and the technical field of immune analysis, in particular to a nanobody for broad-spectrum recognition of Salmonella. Background technique [0002] Salmonella is a foodborne pathogen and a major cause of gastroenteritis, diarrhea and pain. Its infection causes high morbidity and mortality, especially in infants, the elderly, or immunocompromised patients. Three million people die from Salmonella infection every year in the world, among which Salmonella typhimurium serotype and Salmonella enteritidis serotype are the most prevalent. Considering the low infectious doses of most foodborne pathogens, the presence of foodborne pathogens needs to be monitored at every step of food production, processing, distribution and storage. Therefore, the development of rapid and sensitive assays is critical to ensuring the safety of our food supply. [0003] Based on the growth principle of microorganisms, tra...

Claims

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Application Information

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IPC IPC(8): C07K16/12C12N15/13G01N33/569G01N33/543G01N33/535
CPCC07K16/1235C07K16/005G01N33/56916G01N33/54306G01N33/535C07K2317/569C07K2317/565C07K2317/567C07K2317/92C07K2317/94C07K2317/33G01N2333/255G01N2469/10
Inventor 王妍入任亚荣王建龙季艳伟魏娟
Owner NORTHWEST A & F UNIV
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