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High-purity bovine serum albumin purifying process suitable for pilot scale test mass production

A bovine serum albumin, high-purity technology, applied in the field of albumin preparation, can solve the problems of poor utilization of plasma resources, consumption of buffer solution, cumbersome process, etc., and achieve high yield, short process, and process steps little effect

Active Publication Date: 2019-05-21
TIANJIN UNIVERSITY OF TECHNOLOGY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The process is relatively cumbersome, the purity can only reach about 97%, and the plasma resources cannot be well utilized
At present, most of the chromatographic processes for albumin are based on anion exchange and cation exchange, which have a long cycle and consume a lot of buffer.

Method used

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  • High-purity bovine serum albumin purifying process suitable for pilot scale test mass production
  • High-purity bovine serum albumin purifying process suitable for pilot scale test mass production
  • High-purity bovine serum albumin purifying process suitable for pilot scale test mass production

Examples

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Effect test

Embodiment 1

[0033] A purification process for high-purity bovine serum albumin suitable for pilot scale production, comprising the following steps:

[0034] (1) Weigh 8g of finished albumin, dissolve it in 100ml of 20mM NaAc+HAc, and adjust to pH 5.2.

[0035] (2) Preparation of buffer: prepare 4L of equilibration buffer A: 20mMNaAc+HAC, 7L of elution buffer B: 20mMNaAc+HAc, 1L of elution buffer C: 1M NaCl.

[0036] (3) Equilibrium and sample loading: Fill the chromatography column with filler, equilibrate with equilibrium solution A after the chromatography column is installed, flow rate is 20ml / min, after equilibrating about 3 column volumes, start sample loading. Continue to rinse with solution A until the detection line is flat.

[0037] (4) Elution: use solution B for elution, collect the elution peaks and determine the target peaks. After the UV absorption decreased to 50mAU, the peak collection was stopped, and the column was washed with liquid C instead. Finally, wash the colum...

Embodiment 2

[0042] A purification process for high-purity bovine serum albumin suitable for pilot scale production, comprising the following steps:

[0043] (1) Weigh 4g of finished albumin, dissolve it in 40ml of 20mM NaAc+HAc, and adjust to pH 6.

[0044] (2) Preparation of buffer: prepare 3L equilibration buffer A: 20mMNaAc+HA, 3L elution buffer B: 20mMNaAc+HAc, 1L elution buffer C: 1MNaCl

[0045] (3) Equilibration and sample loading: Fill the chromatography column with filler, equilibrate with equilibrium solution A after the chromatography column is installed, flow rate is 12ml / min, after equilibrating about 3 column volumes, start sample loading. Continue to rinse with solution A until the detection line is flat

[0046] (4) Elution: use solution B for elution, collect the elution peaks and determine the target peaks. After the UV absorption decreased to 60mAU, the peak collection was stopped, and the column was washed with liquid C instead. Finally rinse the column with deioniz...

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Abstract

The invention relates to a high-purity bovine serum albumin purifying process suitable for pilot scale test mass production. The method aims at solving the technical problems that an existing bovine serum albumin production process is complex, and the product purity is not high. The one-step chromatography technique is established, and the product with the purity of 99.5% is obtained in the mode that a purchased low-purity bovine serum albumin raw material is subjected to anion exchange chromatography, ultrafiltration and condensation and freeze-drying. The process steps are fewer, the flow path is short, the operation is easy, industrial production is facilitated, and the product purity is high.

Description

technical field [0001] The invention relates to a preparation method of albumin, in particular to a high-purity bovine serum albumin purification process suitable for pilot scale mass production. Background technique [0002] Albumin (Albumin) is a kind of protein that can be dissolved in water and can only be precipitated under high salt concentration, also known as albumin. It is the most abundant protein in serum (or plasma) content, accounting for about 55-63% of its total protein content. Because it is the most abundant and easily purified in vertebrate plasma, it is also the earliest and most clearly studied of all proteins in plasma in the past 30 years. Among them, the amino acid sequences of human serum albumin (human serum albumin, HAS) and bovine serum albumin (bovine serum albumin, BSA) were obtained in 1975. [0003] The industrial production of albumin is still dominated by cold ethanol precipitation. The process is relatively cumbersome, the purity can only...

Claims

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Application Information

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IPC IPC(8): C07K14/765C07K1/18
Inventor 孙永跃何程铖
Owner TIANJIN UNIVERSITY OF TECHNOLOGY