Fluorescent bioprobes for upconversion nanoparticle-labeled aptamers

A fluorescent bioprobe and nanoparticle technology, applied in the field of fluorescent bioprobes, can solve problems such as high background signal, biological nucleic acid damage, potential toxicity, etc., achieve low background signal, avoid damage, and improve detection sensitivity

Inactive Publication Date: 2011-11-30
CHANGCHUN INST OF OPTICS FINE MECHANICS & PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in practical biological applications, QDs need to be excited with short-wavelength excitation light such as ultraviolet light, which will cause damage to the nucleic acid of

Method used

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  • Fluorescent bioprobes for upconversion nanoparticle-labeled aptamers
  • Fluorescent bioprobes for upconversion nanoparticle-labeled aptamers
  • Fluorescent bioprobes for upconversion nanoparticle-labeled aptamers

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specific Embodiment approach 1

[0019] Specific implementation mode one: combine figure 2 To illustrate this embodiment, the fluorescent bioprobe for upconversion nanoparticle-labeled aptamer includes an aptamer labeled with a fluorescent donor, and a DNA strand complementary to the aptamer. The FRET effect occurs as the dye molecule of the fluorescent acceptor, and the UCNPs are used as the fluorescent donor. The UCNPs described in this embodiment are NaYF 4 :Yb, Er, NaYF 4 :Yb, Tm, NaYF 4 :Yb, Ho, LaF 3 :Yb, Er, LaF 3 :Yb, Tm or LaF 3 : one of Yb, Ho, the UCNPs described in the present embodiment are water-soluble, and the UCNPs described in the present embodiment are connected with the aptamer with avidin and biotin.

specific Embodiment approach 2

[0020] Specific implementation mode two: combination figure 2 and image 3 Describe this embodiment, this embodiment is the preparation method of the fluorescent bioprobe of the upconversion nanoparticle-labeled aptamer described in the first embodiment:

[0021] (1) Preparation of UCNPs: 0.272 g of CF 3 COONa, 0.376 g Y(CF 3 COO) 3 , 0.113 g Yb (CF 3 COO) 3 and 0.055g of Er(CF 3 COO) 3 Dissolve it in 10ml of oleylamine, put it into a three-necked bottle, and stir at 100°C for 30 minutes under the protection of argon, then gradually raise the temperature to 320°C and keep it for 1 hour. The product was separated by centrifugation and ultrasonically washed with cyclohexane to obtain a cyclohexane solution of UCNPs.

[0022] (2) Utilize aminophosphoric acid to carry out aqueous phase transfer to the UCNPs prepared in step (1): Weigh 0.13 gram of aminophosphoric acid and disperse it in 5ml ethanol, drop the cyclohexane solution of 3ml UCNPs therein, stir at room temperat...

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Abstract

The invention relates to a fluorescent bioprobe for upconverting nanoparticle-labeled aptamers, which belongs to the technical field of biomolecular detection. In order to overcome the technical problems of existing QDs-labeled aptamer biological probes, including damage to the nucleic acid of the organism where the target molecule is located, high background signal of QDs, potential toxicity, "blinking" effect and chemical instability, provide A fluorescent bioprobe for up-conversion nanoparticle-labeled aptamers, including aptamers labeled with up-conversion nanoparticles as fluorescent donors, and DNA strands complementary to the aptamers, which are connected with fluorescent The donor undergoes a FRET effect as the fluorescent acceptor dye molecule. The fluorescent donors used in the present invention are UCNPs, which have the advantages of low background signal, non-toxicity, no "blinking" effect and strong chemical stability.

Description

technical field [0001] The invention belongs to the technical field of biomolecular detection, in particular, the invention relates to a fluorescent bioprobe labeled with an aptamer by up-converting nanoparticles (UCNPs). Background technique [0002] Aptamers are a special class of nucleotide chains that have high specificity and high affinity for most target molecules, such as organic dye molecules, amino acids, antibiotics, proteins, and even whole cells and microbial pathogens. Aptamers are screened by iterative elution, collection, and reamplification in a synthetic random-sequence oligonucleotide library by means of a phylogenetic process of exponentially enriched ligands. Compared with traditional antibodies, aptamers have many excellent characteristics as biosensing recognition elements. Such as small size, simple preparation process, long shelf life, easy regeneration and chemical modification. The most important of these is that aptamers can fold into variable te...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 刘晓敏孔祥贵涂浪平张友林曾庆辉
Owner CHANGCHUN INST OF OPTICS FINE MECHANICS & PHYSICS CHINESE ACAD OF SCI
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