Directional separation method for immune adjustment intestinal lactobacillus

An immune regulation and directional separation technology, applied in the direction of separation of microorganisms, bacteria, etc., can solve the problems of great differences in activity in the body and increase the complexity of screening work, and achieve the effect of shortening the strain screening cycle and improving the separation selectivity

Active Publication Date: 2017-03-15
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the in vivo activity of many strains with similar in vitro physiological performance is very different [Ibnouzekri et al., 2003], which further increases the complexity of the screening work

Method used

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  • Directional separation method for immune adjustment intestinal lactobacillus
  • Directional separation method for immune adjustment intestinal lactobacillus
  • Directional separation method for immune adjustment intestinal lactobacillus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1: Preparation of streptavidin magnetic beads

[0042] Prepare streptavidin magnetic beads as follows

[0043] (1) Aminated Fe 3 o 4 Preparation of Nanomagnetic Beads

[0044] ① Soak the round-bottom flask, rotor, and reactor liner in aqua regia overnight, then clean them with detergent, then clean them with deionized water and dry them in an oven.

[0045] ②Add 30mL of ethylene glycol to the round bottom flask with rotor, add 6.5g of 1,6-hexanediamine, then add 2.0g of anhydrous sodium acetate, and finally add 1.0g of FeCl 3 ·6H 2 O.

[0046] ③Put the round bottom flask in an oil bath at 50°C with a reflux device, and magnetically stir for about 0.5h-1h. React until the solution in the flask is uniform.

[0047] ④ Carefully transfer the solution in the round-bottomed flask to the liner of the high-pressure reaction kettle lined with tetrafluoroethylene, tighten the reaction kettle, place it at a high temperature of 198°C for 6 hours, and then cool it ...

Embodiment 2

[0058] Example 2: Isolation and screening of immunomodulatory Lactobacillus from human feces using streptavidin magnetic bead technology

[0059] This example uses streptavidin magnetic bead technology to isolate and screen immunomodulatory Lactobacillus from human feces, figure 1 Flowchart for the directed isolation of high-affinity IgA-binding Lactobacillus.

[0060] Specifically include the following steps:

[0061] Isolation and screening of IgA-encapsulated Lactobacillus from feces by streptavidin magnetic beads method:

[0062] ①Weigh 1g of feces from a healthy person, add 10mL of sterilized peptone buffer, and vortex for 5 minutes.

[0063] ②Centrifuge at 1000rpm for 5min, take the supernatant, and filter the large debris and supernatant through a sterile 200-mesh cell strainer into a new tube.

[0064] ③ Centrifuge the filtered centrifuge tube at 10,000 rpm for 5 min, and discard the supernatant. Add 5 mL of peptone buffer, mix by pipetting, centrifuge at 10,000 rp...

Embodiment 3

[0076] Example 3: Isolation and screening of immunomodulatory Lactobacillus from saliva using streptavidin magnetic bead technology

[0077] This example uses streptavidin magnetic bead technology to isolate and screen IgA-encapsulated Lactobacillus from saliva.

[0078] A method for isolating IgA-wrapped Lactobacillus based on streptavidin magnetic bead technology specifically comprises the following steps:

[0079] Preparation of streptavidin magnetic beads: the same as in Example 1.

[0080] Isolation and screening of IgA-encapsulated Lactobacillus from saliva by streptavidin magnetic beads method:

[0081] ① Take 1mL of saliva from a healthy person, add 9mL of sterilized peptone buffer, and vortex for 5 minutes.

[0082] ② After mixing, centrifuge at 10,000 rpm for 5 minutes, and discard the supernatant. Add 5 mL of sterilized peptone buffer, mix by pipetting, centrifuge at 10,000 rpm for 5 min, pour off the supernatant, and repeat again.

[0083] ④Resuspend the precip...

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Abstract

The invention discloses a directional separation method for immune adjustment intestinal lactobacillus, and belongs to the technical field of biological preparations. A biotinylated IgA antibody is used for treating animal or human saliva, intestinal contents or fecal bacterial suspension, streptavidin magnetic nano-beads are used for separation, and enrichment culture and selective separation are performed to obtain the intestinal lactobacillus with an immune adjustment function. Compared with a traditional method, the method can rapidly and directionally separate immune adjustment beneficial bacteria, and obtained strains can be used for feed, common food or functional food after acid resistance, bile resistance and adhesion of the strains are further verified.

Description

technical field [0001] The invention relates to a method for directional isolation of immunoregulatory intestinal lactobacilli, which belongs to the technical field of biological preparations. Background technique [0002] Lactobacillus is an important member of probiotics and widely exists in the gastrointestinal tract of humans and animals. A large number of experiments at home and abroad have confirmed that a variety of lactobacilli in the body can regulate the immune system function of the host, which is why they can prevent infectious diseases, improve food allergies, fight cancer, stabilize the intestinal mucosal barrier, improve the performance of immune adjuvants and relieve intestinal inflammation. The basis of inflammation and other effects. Its mechanism of action is related to its direct activation of immune cells and secretion of immune regulators. [0003] A large number of studies on the isolation of intestinal immunomodulatory Lactobacillus have been carrie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02
CPCC12N1/02C12N1/20
Inventor 孙进朱华玲齐策
Owner JIANGNAN UNIV
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