Bacteria modified by photo-thermal agent and preparation method and application thereof

A technology of photothermal agent and bacteria, which is applied in the field of biomedicine, can solve the problems of limiting tumor target delivery efficiency, etc., and achieve the effect of stable triple photothermal heating effect, strong penetration ability and long-lasting retention effect

Pending Publication Date: 2022-05-27
RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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Problems solved by technology

In addition, indocyanine green (ICG) is a near-infrared fluorescent dye widely used in the medical field, which has excellent photothermal conversion ability, but due to its strong lipophilicity and serum protein affinity, free ICG will be quickly cleared by the liver and gallbladder (plasma Half-life <5min), which severely limits its tumor-targeted delivery efficiency

Method used

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  • Bacteria modified by photo-thermal agent and preparation method and application thereof
  • Bacteria modified by photo-thermal agent and preparation method and application thereof
  • Bacteria modified by photo-thermal agent and preparation method and application thereof

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Embodiment

[0107] 1. Preparation of photothermal agent-modified bacteria and testing their performance

[0108] Bacteria were engineered by genetic engineering and surface engineering to construct a ternary photosensitive system containing melanin, ICG, and polydopamine ( figure 1 .a).

[0109] For the melanin-producing BL21mel bacteria (Mel), we transformed the plasmid encoding tyrosinase (pET-28a-melA) into Escherichia coli BL21, containing kanamycin, tryptone, casein, tyrosine and CuSO4 in solid medium, cultured at 30°C for 48h to express melanin ( figure 1 .b). Surface-bound polydopamine-bound bacteria (Mel-pDA) were prepared by oxidation and self-polymerization of dopamine, specifically, Mel (2 × 10 8 CFU) was added to 10 mM tris-HCl buffer containing 0.25 mg / ml dopamine and the mixture was vortexed at 1500 rpm for 15 min. Then, 50 μl of 1 mg / ml ICG in water was added and vortexed for 30 min to bind to pDA by hydrogen bonding, π-π stacking and ionic bonding. Finally, washed twi...

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Abstract

The invention belongs to the field of biological medicine, and relates to a bacterium modified by a photo-thermal agent, and a preparation method and application thereof. The bacterium is an engineered bacterium combining photothermal agent surface modification and genetic expression, and is a bacterium which expresses melanin in a cell and is modified by at least one photothermal agent on the outer surface of the cell. The at least one photo-thermal agent comprises a first photo-thermal agent and/or a second photo-thermal agent, the first photo-thermal agent is dopamine, and the second photo-thermal agent comprises one or more selected from photo-thermal dyes; preferably, the second photo-thermal agent is a near-infrared fluorescent dye, and more preferably, indocyanine green. The bacterium modified by the photothermal agent has a stable triple photothermal heating effect after laser irradiation, has specific colonization capacity at a tumor part, is high in infiltration capacity, and can realize uniform distribution and a lasting retention effect.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a bacterium modified by a photothermal agent, a preparation method and application thereof. The bacterium is an engineered bacterium that combines surface modification of a photothermal agent and genetic expression, specifically, a bacterium that expresses melanin in cells and is modified by at least one photothermal agent on the extracellular surface. The photothermal agent-modified bacteria have stable triple photothermal heating effect after laser irradiation, have specific colonization ability at tumor sites, and have strong penetration ability, and can achieve uniform distribution and long-lasting retention effect. Background technique [0002] Photothermal therapy (PTT) is a local treatment method that directly acts on the lesion site. It ablates solid tumors through hyperthermia generated by photosensitizers under electromagnetic radiation. It has good clinical application and tra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/00A61K41/00A61K47/51A61P35/00
CPCC12N1/20C12N1/005A61K41/0052A61K47/51A61P35/00A61K2300/00Y02A50/30
Inventor 刘尽尧郭海燕
Owner RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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