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A low-cost preparation method of igf-1 and its application

A technology of IGF-1 and ELP-IGF-1, which is applied in the field of protein engineering, can solve the problems of time-consuming and labor-consuming, high price of chromatographic separation packing materials, and difficult scale-up of chromatographic separation methods, and achieve simplified preparation procedures and low-cost preparation Protocol, Effect of Efficient Soluble Expression

Active Publication Date: 2021-10-29
苏州浦泉生物科技有限公司
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  • Claims
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Problems solved by technology

The chromatographic separation method requires multiple steps such as sample loading, binding, washing, and elution, which is time-consuming and labor-intensive. In addition, it is difficult to scale up the chromatographic separation method in industrial applications, and the price of the filler required for the chromatographic separation method is high.

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  • A low-cost preparation method of igf-1 and its application
  • A low-cost preparation method of igf-1 and its application

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Embodiment Construction

[0027] The specific embodiments of the present invention will be further described below in conjunction with the accompanying drawings. It should be noted here that the descriptions of these embodiments are used to help understand the present invention, but are not intended to limit the present invention. In addition, the technical features involved in the various embodiments of the present invention described below may be combined with each other as long as they do not constitute a conflict with each other.

[0028] In addition, the terms "first" and "second" are used for descriptive purposes only, and cannot be interpreted as indicating or implying relative importance or implicitly specifying the quantity of indicated technical features. Thus, the features defined as "first" and "second" may explicitly or implicitly include at least one of these features. In the description of the present invention, "plurality" means at least two, such as two, three, etc., unless otherwise ...

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Abstract

The invention discloses a low-cost preparation method of IGF-1, comprising: S1: construction of ELP-IGF-1 plasmid; S2: construction of engineering bacteria; S3: cultivating the above-mentioned engineering bacteria in TB medium, inducing overnight, and collecting Bacteria, ultrasonically disrupted, and centrifuged at low temperature to obtain the supernatant; S4: heat the supernatant obtained in the above steps in a water bath, and centrifuge to obtain the precipitate; S5: dissolve the precipitate with pre-cooled Tris-HCl buffer, and centrifuge to obtain the supernatant; S6 : Overnight enzyme digestion; S7: After the enzyme digestion is completed, heat the protein solution in a water bath at 30°C, centrifuge at 30°C and take the supernatant, use magnetic beads to bind TEV enzyme to the supernatant, collect the beads by centrifugation, and freeze-dry the supernatant This is followed by IGF‑1. IGF‑1 is used to promote cell growth and proliferation. The invention provides a brand-new preparation method without any chromatographic filler, and the prepared protein should be used to stimulate the growth and proliferation of cells containing IGF-1 receptors.

Description

technical field [0001] The invention belongs to the technical field of protein engineering, and in particular relates to a low-cost preparation method of IGF-1 and its application. Background technique [0002] Insulin-like growth factor-1 (insulin like growth factor, IGF-1) is one of the most important growth factors on the growth hormone IGF axis. IGF-1 is a single polypeptide protein consisting of 70 amino acids with a molecular weight of 7649 Da. IGF-1 is an anabolic hormone produced in the liver that stimulates the proliferation and differentiation of various cell types and plays an important role in tissue renewal and repair. IGF-1 can regulate the growth of somatic cells, and studies have shown that it is involved in various physiological processes in mammals. In addition, related studies have also shown that IGF-1 plays a vital role in cell energy metabolism, growth and development. Based on IGF-1 participating in various important metabolic processes of cells, IG...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/65C12N15/70C12N15/16C12N15/66
CPCC07K14/65C12N15/66C12N15/70
Inventor 黄凯宗汪舒姝
Owner 苏州浦泉生物科技有限公司
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