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Preparation of novel aggregation-induced fluorescent polypeptide probe for detecting bacterial endotoxin and application

A technology of aggregation-induced fluorescence and bacterial endotoxin, applied in fluorescence/phosphorescence, luminescent materials, biological testing, etc., to achieve the effects of mature technology, short acquisition period and short detection time

Inactive Publication Date: 2019-07-23
NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the rapid detection of bacterial endotoxins has not been reported

Method used

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  • Preparation of novel aggregation-induced fluorescent polypeptide probe for detecting bacterial endotoxin and application
  • Preparation of novel aggregation-induced fluorescent polypeptide probe for detecting bacterial endotoxin and application
  • Preparation of novel aggregation-induced fluorescent polypeptide probe for detecting bacterial endotoxin and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027]Example 1: Preparation of aggregation-induced fluorescent polypeptide probe TPE-pep for detection of bacterial endotoxin

[0028] Experimental material: polypeptide whose sequence is: GCKPTFRRLKWKYKCG, Sangon Bioengineering (Shanghai) Co., Ltd.; 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), batch number 090M14531V, Shanghai Aladdin Reagent Co., Ltd.; Succinimide (NHS), batch number MKBG7914V, Shanghai Aladdin Reagent Co., Ltd.;

[0029] Experimental instruments and equipment: high performance liquid chromatography, Agilent Corporation of the United States.

[0030] experiment procedure:

[0031] Dissolve 6 mg of the aggregation-inducing fluorescent compound 1-(4-carboxyphenyl)-1,2,2-triphenylethylene in the solvent dimethyl sulfoxide, 11 mg 1-(3-dimethylaminopropyl)-3-ethyl Add 7 mg of carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) to the above solution, shake at room temperature for 30 to 60 minutes, then add 8 mg of the polypeptid...

Embodiment 2

[0032] Example 2: TPE-pep detects different concentrations of LPS in solution

[0033] Experimental materials: bacterial endotoxin lipopolysaccharide (LPS), purchased from Sigma Co., Ltd., USA; HEPES buffer, purchased from Sangon Bioengineering (Shanghai) Co., Ltd.; fluorescent 96-well plate, purchased from Corning Co., Ltd., USA; The glassware was fully cleaned with ultrapure water, and then sterilized at high temperature.

[0034] Experimental instruments and equipment: AY120 electronic analytical balance; TCS10 constant temperature mixer; TECAN SPARK 10M multifunctional microplate reader; Guanghao ZF-20D ultraviolet dark box.

[0035] Experimental process: Accurately weigh LPS plus HEPE buffer solution to make it into a standard solution with a concentration of 400μg / ml (40μM), and then dilute it according to the concentration gradient to 4μM, 3μM, 2μM, 1.4μM, 0.4μM, 0.3μM, 0.2μM 11 concentrations of μM, 0.14 μM, 0.04 μM, 0.03 μM and 0.01 μM. Each dilution needs to be ful...

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Abstract

The invention belongs to the field of biotechnology and clinical medical diagnosis and relates to preparation of a novel aggregation-induced fluorescent polypeptide probe for detecting bacterial endotoxin and application. On the basis of aggregation-induced emission effect (AIE), polypeptide is modified on the structure of the probe, and linear fluorescence enhancement is generated by utilizing specific combination with the bacterial endotoxin, so that the concentration of the bacterial endotoxin is detected. The preparation and the application have the advantages of simple operation, fast andaccurate detection, strong specificity and high sensitivity and the like, and have good application prospect.

Description

technical field [0001] The invention belongs to the fields of biotechnology and clinical medical diagnosis, and relates to the preparation and application of a novel aggregation-induced fluorescent polypeptide probe for detecting bacterial endotoxin. Based on the aggregation-induced fluorescence effect (AIE) molecule, a polypeptide is modified on its structure, and its specific binding to bacterial endotoxin is used to generate linear fluorescence enhancement, thereby detecting the concentration of bacterial endotoxin. Background technique [0002] Bacterial endotoxins are characteristic structures of the cell walls of Gram-negative bacteria that are released when the bacteria die, disintegrate, or have their cell walls broken down. Its chemical composition includes lipid A, core polysaccharide and O-specific antigen, wherein lipid A is the active component of bacterial endotoxin toxicity, core polysaccharide has a constant structure in different strains, and O-specific anti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C09K11/06G01N21/64G01N33/569G01N33/58
CPCC09K11/06G01N21/6428G01N33/56911G01N33/582C09K2211/10
Inventor 朱栋唐莹莹杨小彤
Owner NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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