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Platelet growth factor and extraction method and application thereof

An extraction method and growth factor technology, applied in the biological field, can solve problems such as complex steps and protein denaturation

Pending Publication Date: 2021-02-09
INFINITY SGC HK LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The steps of traditional technology are complicated, and the protein is easily denatured by ultrasound and repeated freezing and thawing

Method used

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  • Platelet growth factor and extraction method and application thereof
  • Platelet growth factor and extraction method and application thereof
  • Platelet growth factor and extraction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052]The blood for preparing the PRP and SGC samples in the examples of the present invention were all from the Hong Kong Red Cross.figure 1 Shows the preparation process of SGC samples. SGC preparation and combing consists of three steps:

[0053]First: Preparation of Platelet-rich Plasma (PRP)

[0054]This step adopts a stepped centrifugation method, specifically through two steps of centrifugation at different speeds, the centrifugation conditions are 1000 rpm for 10 minutes, 3000 rpm for 15 minutes, and the supernatant is extracted to obtain PRP.

[0055]The magnetic seat method is used to remove iron ions in PRP.

[0056]Second: Freeze and Thaw

[0057]The product PRP obtained in the first step is frozen and thawed, platelets are burst, and cytokines and growth factors are released to form a growth group.

[0058]Freeze-thaw is to freeze platelet-rich plasma PRP at -80°C for 10 hours, then take it out and place it in a 37°C water bath to immediately thaw for 10 minutes, save the sample and obse...

Embodiment 2

[0068]Example 2 Preparation of SGC, an optimization technique for autologous growth groups

[0069]This embodiment exemplarily provides a preparation method for optimizing the autologous growth group SGC, wherein the blood is derived from humans. Specifically, the experiment carried out includes the following steps:

[0070]1. Separate blood. A 50ml blood sample was separated by two steps of centrifugation at different speeds (centrifugation conditions were 1250 rpm for 5 minutes and 3800 rpm for 20 minutes) to obtain platelet-rich plasma (PRP).

[0071]2. Use the magnetic seat method to remove the iron ions in the PRP obtained in step 1.

[0072]3. Lysate high-concentration platelet plasma by freezing and thawing; wherein the freezing and thawing conditions are as follows: freeze PRP at -80°C for 10 hours, and quickly thaw at 37°C for 10 minutes to rupture the platelets in the PRP obtained in step 2. To obtain isolates containing high concentrations of cytokines and growth factors, including: ...

Embodiment 3

[0077]In order to standardize SGC products, this example uses an ELISA kit to detect platelet factor 4 (PF4), β-thromboglobulin (β-TG), PRP prepared according to the method of Example 2 and the optimized autologous growth group SGC. Epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF) content. Simultaneously compare the growth factors in PRP, SGC and whole blood (WB).

[0078]Specifically, the experiment carried out includes the following steps:

[0079]1. Using the PRP, SGC prepared in Example 2 and homologous whole blood (WB) without any treatment.

[0080]2. Using Human PF4, β-TG, EGF, VEGF, PDGF ELISA kits, add PRP, SGC and WB to different ELISA kits to coat human PF4, β-TG, EGF, VEGF, In a 96-well plate of PDGF antibody, add 50μL per well, make 3 replicate wells for each sample, and incubate for 30 minutes at room temperature.

[0081]3. Add the mixture containing the capture antibody and detection antibody to a 96-well plate, 50 μ...

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Abstract

The invention provides a platelet growth factor as well as an extraction method and application thereof. The extraction method of the platelet growth factor comprises the step of directly freezing andthawing platelets. Through long-term research, the inventor finds that the growth factor can be fully released by directly freezing and thawing the platelets once, and the activity of the growth factor is relatively high, i.e., effective extraction of the high-activity growth factor is realized through simple steps. The platelet growth factor extracted by the method can be used for autologous skin wound healing, skin regeneration, skin whitening or skin oxidation / aging resistance.

Description

Technical field[0001]The invention belongs to the field of biotechnology, and specifically relates to a platelet growth factor and an extraction method and application thereof.Background technique[0002]Growth factors are proteins naturally present in the body and have the ability to stimulate and enhance cell growth and tissue repair. Platelets in the blood store a large number of active growth factors. Upon activation, these growth factors are released in large quantities, which play an extremely important role in promoting wound healing, cell proliferation and differentiation, and tissue formation. Based on this, platelet rich plasma (PRP, Platelet Rich Plasma) is more and more widely used in clinics, including orthopedics, ophthalmology, and plastic surgery.[0003]However, PRP extracted from patients must be fresh and cannot be stored for a long time. The average lifespan of platelets is usually only 5 to 9 days after extraction, but normal PRP treatment procedures need to be perf...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/475C07K14/49C07K14/485C07K1/14A61K38/18A61P17/02A61K8/64A61Q19/02A61Q19/08
CPCA61K8/64A61K38/1858A61Q19/02A61Q19/08A61P17/02C07K14/475C07K14/485C07K14/49
Inventor 詹华强蔡慧思蔡慧琪陈嘉伦董婷霞
Owner INFINITY SGC HK LTD
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