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Staphylococcus aureus staphyloccucal enterotoxins B resistant monoclonal antibody YG11-2 and application thereof

An antibody and sequence listing technology, applied in applications, antibodies, antibacterial drugs, etc., can solve the problem of SEB poisoning vaccines and drugs, etc., and achieve the effect of high affinity and improving survival rate.

Active Publication Date: 2020-06-26
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no effective vaccine and drug against SEB poisoning clinically

Method used

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  • Staphylococcus aureus staphyloccucal enterotoxins B resistant monoclonal antibody YG11-2 and application thereof
  • Staphylococcus aureus staphyloccucal enterotoxins B resistant monoclonal antibody YG11-2 and application thereof
  • Staphylococcus aureus staphyloccucal enterotoxins B resistant monoclonal antibody YG11-2 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1, the preparation of His-SEB protein and its functional research

[0040] 1. Preparation of His-SEB protein

[0041] 1. Replace the small fragment between the NdeI and XhoI restriction sites of the vector pET28a(+) with the DNA molecule shown in sequence 6 of the sequence listing to obtain the recombinant plasmid pET28a-SEB. Sequencing verified.

[0042] In the recombinant plasmid pET28a-SEB, the DNA molecule shown in sequence 6 in the sequence table is fused with the DNA molecule encoding His tag on the vector backbone to form a fusion gene to express the fusion protein.

[0043] The fusion gene is composed of the following two segments sequentially from upstream to downstream: the DNA molecule shown in sequence 8 in the sequence listing, and the DNA molecule shown in sequence 6 in the sequence listing.

[0044] The fusion protein consists of the following two segments sequentially from the N-terminal to the C-terminal: the segment shown in sequence 7 of ...

Embodiment 2

[0073] Embodiment 2, phage antibody library screening anti-SEB protein monoclonal antibody

[0074] 1. Biopanning of anti-SEB monoclonal antibodies

[0075] (1) SEB group

[0076] 1. The first round of affinity panning

[0077] (1) Coat the immunotube with 500 μL of His-SEB protein solution (adjust the protein concentration with PBS buffer) with a protein concentration of 10 μg / mL, and overnight at 4°C.

[0078] (2) Take the immunotube, add PBST buffer solution containing 5g / 100mL skimmed milk powder, and block at room temperature for 1 hour.

[0079] (3) Take the phage antibody library, add PBST buffer containing 5g / 100mL skimmed milk powder, and block at room temperature for 1 hour.

[0080] (4) Take the immune tube that completed step (2), first add sterile PBS buffer to wash 3 times, and then add the phage antibody library that completed step (3) (the input amount of phage is about 1.2×10 12 ), let stand at room temperature for 1h.

[0081] (5) After completing step (...

Embodiment 3

[0101] Example 3, Functional Identification of YG11-2 Antibody

[0102] 1. Preparation of YG11-2 antibody

[0103] 1. Construction of recombinant plasmids

[0104]The small fragment between the HindIII and XhoI restriction sites in the vector pCDNA3.1(+) was replaced with the DNA molecule shown in Sequence 2 of the sequence listing to obtain the heavy chain expression vector. In sequence 2 of the sequence listing, nucleotides 1-584 constitute a promoter, nucleotides 681-2108 encode a full-length heavy chain, and nucleotides 2141-2196 constitute a terminator.

[0105] The small fragment between the HindIII and XhoI restriction sites in the vector pCDNA3.1(+) was replaced with the DNA molecule shown in Sequence 4 of the sequence listing to obtain the light chain expression vector. In sequence 4 of the sequence listing, nucleotides 1-584 constitute a promoter, nucleotides 681-1388 encode a full-length light chain, and nucleotides 1421-1476 constitute a terminator.

[0106] 2. ...

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Abstract

The invention discloses a staphylococcus aureus staphyloccucal enterotoxins B resistant monoclonal antibody YG11-2 and application thereof. The invention protects an IgG antibody (named as YG11-2 antibody) which consists of a light chain and a heavy chain, wherein a heavy chain variable region in the heavy chain consists of a sequence 1 in a sequence table from a 21-145th amino acid residue at a tail end N; and a light chain variable region in the light chain consists of a sequence 3 in a sequence table from a 21-128th amino acid residue at a tail end N. The invention further protects application of the YG11-2 antibody in preparing a medicine for treating poisoning of staphyloccucal enterotoxins B, application in preparing a medicine for preventing the staphyloccucal enterotoxins B, and application in preparing a medicine for preventing and / or treating diseases caused by staphylococcus aureus. The monoclonal antibody YG11-2 has significant application prospects in treating poisoning ofthe staphyloccucal enterotoxins B and preventing or treating diseases caused by the staphylococcus aureus.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a monoclonal antibody YG11-2 against Staphylococcus aureus enterotoxin B and its application, in particular to the preparation of the monoclonal antibody for preventing and / or treating Staphylococcus aureus-induced application in medicine for diseases. Background technique [0002] Staphylococcus aureus (Staphylococcus aureus) is a Gram-positive pathogenic bacteria widely distributed in nature. It is estimated that 20-30% of the population carries the pathogen, which mainly exists in the mucous membranes, skin, and especially the nasopharynx of the human body. Staphylococcus aureus can cause a range of diseases, from minor skin infections, to scalded skin syndrome, to abscesses, to life-threatening pneumonia, meningitis, endocarditis, osteomyelitis, toxic shock syndrome, etc. . Staphylococcus aureus infection affects a wide range of skin, soft tissue, respiratory tract,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/12C12N15/13A61K39/40A61P31/04
CPCC07K16/1271A61P31/04C07K2317/92A61K2039/505
Inventor 杨光刘成华刘玉刘方杰高亚萍冯健男
Owner ACADEMY OF MILITARY MEDICAL SCI
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