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A kind of luteinizing hormone analogue and preparation method thereof

A technology for luteinizing hormone and analogs, applied in the field of veterinary luteinizing hormone analogs and their preparation, to achieve the effects of superior practicability and commercial value

Active Publication Date: 2021-04-27
HUAQIAO UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The invention provides a luteinizing hormone analogue and a preparation method thereof. Aiming at the existing technical problems, the present invention designs a suitable linker sequence according to the length, foldability, hydrophobicity and charge effect of the linker sequence so as to obtain different time-effective drugs. A series of veterinary luteinizing hormone analogues

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  • A kind of luteinizing hormone analogue and preparation method thereof
  • A kind of luteinizing hormone analogue and preparation method thereof
  • A kind of luteinizing hormone analogue and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Example 1, Preparation of Luteinizing Hormone Analogs of Different Glycosylation Types

[0039] Using bovine (bovine, b) LHα and LHβ gene sequences as templates, according to the relationship between the structure and function of luteinizing hormone, the two subunits of LH were connected in the order of NH2-β-linker–α-COOH to form a single-chain fusion LH. According to structural characteristics, such as amino acid sequence length, foldability, hydrophobicity, charge effect and glycosylation degree, 10 different linker sequences were designed and connected with wild-type and codon-biased bLHα and bLHβ respectively to form 20 fusion proteins with different sequences.

[0040] 1.1 Design of recombinant bLH single-chain fusion protein

[0041] Obtain the bovine (bovine) LHα subunit coding sequence (NM_173901.3) and LHβ subunit coding sequence (M11506.1) from NCBI's gene bank Genebank, which will carry the start codon and signal peptide coding sequence but delete the stop...

Embodiment 2

[0150] Example 2, Determination of Relative Biological Activity of Recombinant LH Carrying Different Linker Sequences

[0151] The recombinant bLH carrying 0, 1 and 2 N-glycosylation sites and O-glycosylation sites in the linker sequence was tested for biological activity according to the Luteinizing Hormone Bioassay in Pharmacopoeia.

[0152] 2.1 Preparation of experimental materials

[0153] 2.1.1 pcDNA3.1(+) transfected CHO-K1 cell culture supernatant and bLH transfected cell supernatant carrying different linkers (see Table 2.1).

[0154] Table 2.1 Properties of each bLH protein sample

[0155]

[0156] 2.1.2 Preparation of 10× solvent

[0157] 500mgBSA, 4.5gNaCl, add distilled water to 40mL, adjust the pH to 7.2 with sodium hydroxide, set the volume to 50mL, shake and mix.

[0158] 2.1.3 Configuration of standard injection:

[0159] On the day of the experiment, take the standard substance of urogonadotropin, cut open the ampoule, wash out the dry powder of the sta...

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Abstract

The invention discloses a luteinizing hormone (luteinizing hormone, LH) analogue and a preparation method thereof. The luteinizing hormone analogue is connected from N-terminus to C-terminus in the order of LHβ-linker sequence-LHα, designed The linker sequences have different lengths, foldability, hydrophobicity, different numbers and types of glycosylation sites and histidine tags, and are fused with the α and β subunits of natural luteinizing hormone to form different time-sensitive drugs. Functional single-chain polypeptide, which has superior practicability than natural luteinizing hormone.

Description

【Technical field】 [0001] The invention belongs to the field of biotechnology, in particular to a veterinary luteinizing hormone analog (luteinizing hormone, LH) and a preparation method thereof. 【Background technique】 [0002] Luteinizing hormone (LH) is a heterodimeric glycoprotein secreted by basophils of the anterior pituitary gland, which is formed by the combination of two subunits, LHα and LHβ, in a non-covalent manner. LH and follicle-stimulating hormone (FSH) play a synergistic role in the regulation of the functions of the ovaries of female organisms and the testes of male organisms. FSH mainly promotes follicular development. LH mainly promotes follicle maturation and ovulation, and at the same time promotes corpus luteum production. During the reproductive cycle of female animals, the concentration of LH and FSH in the blood have different cycle change curves. A brief LH peak is required at ovulation, and a low level of LH is required throughout the reproductiv...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/85
CPCC07K14/59C07K2319/21C12N15/85C12N2800/22
Inventor 林俊生刘军伟刁勇
Owner HUAQIAO UNIVERSITY
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