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Process for separating megakaryocyte stimulating factor from human plasma and purifying it and its application

A technology of megakaryocytes and stimulators, applied in the field of biochemical separation and purification, can solve the problems of low activity, large volume, and inconvenient production

Inactive Publication Date: 2002-09-18
NANJING UNIV
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  • Summary
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AI Technical Summary

Problems solved by technology

In view of the limited source of fetal kidney, although human urine is unlimited, it is bulky and must be enriched from a large amount of urine, which brings inconvenience to production. Moreover, human urine MES is mainly in the form of low molecular weight, and its activity is much lower than that of high molecular weight form.

Method used

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Examples

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Effect test

Embodiment

[0029] Example: Collect 300g wet weight of plasma precipitate after preparing superalbumin (equivalent to 100L plasma), dissolve in 6L 10mmol / L PBS of pH6.5, centrifuge at 4°C, 9000r / m for 10 minutes, and take the supernatant (Protein concentration 2.46mg / ml), obtain protein 14.76g altogether, with centrifugal supernatant upper DEAE cellulose column chromatography, with 10mmol / L PBS and the 10mmol / L PBS that contains 0.1mol / L NaCl to wash away impurity protein, again A total of 10.5 g of the target protein was washed with 0.5 mol / L NaCl / 10 mmol / L PBS, and the recovery rate was 71.1%. join (NH 4 ) 2 SO 4 To 80% saturation, the precipitate was collected and dialyzed against water; the dialysate was separated by a Sephacryl gel column, and a total of 4 protein peaks were obtained, and peak I and peak II were collected to obtain 4.29 g of protein with a recovery rate of 40%. After freeze concentration, Balanced by dialysis, separated by DEAE-Sephadex column. After loading the s...

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PUM

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Abstract

A process for separating megakarylcyte stimulator (MES) from human plasma and purifying it includes dissolving the deposite of human plasma, which is generated by extracting albumin and globulin centrifugal treating, ultrafiltrating, ion-exchange cellulose chromatography, gel filter and DEAE gel chromatography. The said MES has 40-42 Kda or 18-20 kda of molecular weight, and can promote the generation of thrombocytes and the multiplication of naval blood's stem cells, so it can be used for treating thrombocytopenia and stem cell transplant.

Description

1. Technical field: [0001] The invention belongs to the technical field of biochemical separation and purification. 2. Background technology [0002] Thrombocytopenia can be caused by many factors, such as exposure to harmful chemical substances, viral infection, radiation, chemotherapy, and certain blood diseases (such as thrombocytopenic purpura, myelodysplastic syndrome-refractory anemia, aplastic anemia) . The consequences of low platelets are serious, causing dangerous bleeding that can appear as large purpura on the skin, and bleeding into the internal organs or brain can be life-threatening. At present, there is no effective platelet-increasing drug on the market. In 1994, three companies in the United States synthesized the large N-terminal fragment of thrombopoietin (Thrombopoietin, TPO) through DNA recombination technology. Hope, but the clinical effect of phase I and II is not satisfactory, so it has not been marketed so far. Other factors such as IL-3, 6, 11 ar...

Claims

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Application Information

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IPC IPC(8): A61K38/18A61P7/06C07K14/475C12N5/074
Inventor 张鹤云金以丰张太平汤国枝袁达文
Owner NANJING UNIV
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