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Apolipoprotein B nano antibody and coding sequence and application thereof

A technology of apolipoprotein and nano-antibody, which is applied in the field of biomedicine or biopharmaceuticals, and can solve the problems of low sensitivity, poor antibody stability, and high production cost

Inactive Publication Date: 2013-10-09
SOUTHEAST UNIV
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AI Technical Summary

Problems solved by technology

[0003] The working principle of the kit (immunoturbidimetric method) for detecting apolipoprotein B currently on the market is mainly realized by an antibody against apolipoprotein B, but this antibody in the traditional sense has poor stability and low sensitivity. Low and high production costs, all of which limit the detection of apolipoprotein B

Method used

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  • Apolipoprotein B nano antibody and coding sequence and application thereof
  • Apolipoprotein B nano antibody and coding sequence and application thereof
  • Apolipoprotein B nano antibody and coding sequence and application thereof

Examples

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Effect test

Embodiment 1

[0025] Example 1: Construction of a Nanobody Library against Apolipoprotein B:

[0026] (1) The purity of apolipoprotein B extracted from human blood (purchased from Nanjing Huibiao Biotechnology Co., Ltd.) was detected by SDS-PAGE electrophoresis gel, figure 1 The protein bands from left to right are: the first is the protein molecular standard, and the second is the sample apolipoprotein B; figure 1 It shows that the purity of apolipoprotein B has reached more than 99%, and the concentration of apolipoprotein B used for immunization is 500 micrograms per milliliter. For each immunization, 20 mg of apolipoprotein B is mixed with an equal volume of Freund's adjuvant. A Xinjiang Bactrian camel (Jurong Shenglong Livestock Breeding Factory) was immunized once a week for a total of 4 times. Except for the first use of complete Freund's adjuvant, the remaining times were all used Freund's incomplete adjuvant. During the immunization process B cells are stimulated to express antige...

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Abstract

The invention discloses a VHH chain of a nano antibody of apolipoprotein B, which comprises a framework region (FR) and a complementary determining region (CDR). The invention discloses an amino acid sequence of the FR selected from the lower group and an amino acid sequence of the CDR. The invention also discloses an apolipoprotein B nano antibody and a DNA molecule for coding the VHH chain of the nano antibody of apolipoprotein B or the apolipoprotein B nano antibody, and discloses a host cell capable of expressing the nano antibody of apolipoprotein B. Moreover, the invention discloses an application of the apolipoprotein B nano antibody for detecting the apolipoprotein B. Through the nano antibody gene sequence and host cell disclosed by the invention, the nano antibody can be efficiently expressed in Escherichia coli and is used for developing an apolipoprotein B detection reagent.

Description

technical field [0001] The invention belongs to the technical field of biomedicine or biopharmaceuticals, and relates to a nanobody directed at apolipoprotein B. Background technique [0002] Apolipoprotein (apolipoprotein, apo) is a protein in plasma lipoprotein, and its basic function is to carry lipids. Apolipoprotein B (apoB) exists on the surface of low-density lipoprotein, and the recognition and uptake of LDL by cells is mainly realized by recognizing apoB. Due to differences in amino acid composition, apoB can be divided into the following subclasses: apoB48 and apoB100. apoB48 is one of the apolipoproteins of chylomicrons (CM); apoB100 is one of the apolipoproteins of very low-density lipoprotein (VLDL) and low-density lipoprotein (LDL). When apolipoprotein B increases, even if the LDL level is normal, the incidence of coronary heart disease can also be increased. ApoB is the main protein of LDL (low-density lipoprotein), therefore, apoB in serum mainly represent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C12N15/13C12N15/70C12N1/21G01N33/68
Inventor 万亚坤孙燕燕母亚雯谢维
Owner SOUTHEAST UNIV
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