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Nucleic acid aptamer for combining human beta-microglobulin

A nucleic acid aptamer and microglobulin technology, which is applied in DNA preparation, DNA/RNA fragments, recombinant DNA technology, etc., can solve problems such as iron overload and hemochromatosis

Inactive Publication Date: 2014-03-26
杭州耀洲生物科技有限公司 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, together with the HFE protein, it participates in the regulation of pinocytosis of iron into the small intestine, and the default of this function can lead to iron excess and hemochromatosis

Method used

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  • Nucleic acid aptamer for combining human beta-microglobulin
  • Nucleic acid aptamer for combining human beta-microglobulin
  • Nucleic acid aptamer for combining human beta-microglobulin

Examples

Experimental program
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Effect test

Embodiment Construction

[0069] (a) 200ug beta2-microglobulin was dissolved in 200ul water.

[0070] (b) Take 0.08g of Epoxy-activated Sepharose, add 1ml of water to make it fully swell, after about 5 minutes, after the distribution is uniform, centrifuge at low speed, and remove the supernatant. Wash several times to remove additives.

[0071] (c) Take the beta2-microglobulin from step (a), add it to the washed Epoxy-activated Sepharose, and then add 1M NaHCO 3 , reacted at room temperature for 16 hours.

[0072] 2. Block the active group of unbound beta2-microglobulin:

[0073] (1) Reaction product, with 0.1M NaHCO 3 Wash three times.

[0074] (2) The coupled product was placed in 1M ethanolamine, and reacted at 40°C for 4 hours.

[0075] (3) Wash three times alternately with 0.1M NaAc-0.5M NaCl solution and 0.1M Tris-HCL (containing 0.5M NaCl, pH8.0) solution.

[0076] (4) Finally, wash twice with 10% ethanol, and store the product in 10% ethanol.

[0077] 3. SELEX process

[0078] (1) Take 8...

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Abstract

The invention discloses a short-chain nucleotide aptamer with high specificity and high affinity of beta-microglobulin and a screening method of the aptamer. The nucleotide aptamer provided by the invention comprises DNA (deoxyribonucleic acid) segments of nucleic acids shown by any one from a sequence 1 to a sequence 10. The preparation method of the nucleotide aptamer comprises the steps of designing and synthesizing a random single-chain nucleotide library, and screening the nucleotide aptamer of the beta-microglobulin. The nucleic acid aptamer disclosed by the invention is used for identifying a combined target, so that the nucleic acid aptamer is favorable for rapid determination of the beta-microglobulin.

Description

technical field [0001] The present invention generally relates to nucleic acid aptamers, and more specifically relates to nucleic acid aptamers combined with human Beta2-microglobulin and a screening method thereof. Background technique [0002] Beta2-microglobulin widely exists in blood, spinal fluid, saliva and urine, and is the main component of the major histocompatibility antigen MHC-I on the cell surface. MHC-I molecules exist on the surface of almost all cells except red blood cells [1，2] . Determination of beta2-microglobulin content in serum or plasma is helpful to clinically determine the biological activity of the cellular immune system and the activity of tumor markers. [0003] The molecular weight of beta-microglobulin is 12,000. It is a spherical structure composed of multiple beta-sheet domains. It does not have a membrane-passing domain. It is one of the four subunits that constitute MHC-I molecules. It plays a role in maintaining the structure of the MHC-...

Claims

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Application Information

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IPC IPC(8): C12N15/115C12N15/10
Inventor 贾俊玲王晓庆张莹莹冯安娜王铄余盛朱勤峰朱德领郭锰
Owner 杭州耀洲生物科技有限公司
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