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A high-purity bovine serum albumin purification process suitable for pilot scale production

A bovine serum albumin, high-purity technology, applied in the field of albumin preparation, can solve the problems of long cycle, buffer consumption, cumbersome process, etc., and achieve the effect of simple operation, short process and few process steps

Active Publication Date: 2022-03-29
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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  • A high-purity bovine serum albumin purification process suitable for pilot scale production
  • A high-purity bovine serum albumin purification process suitable for pilot scale production
  • A high-purity bovine serum albumin purification process suitable for pilot scale production

Examples

Experimental program
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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) Equilibrium 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 deionized...

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Abstract

The invention relates to a method for purifying high-purity bovine serum albumin suitable for pilot-scale mass production, and aims to solve the technical problems that the existing bovine serum albumin production process is relatively complicated and the product purity is not high. The invention establishes a one-step chromatographic technique, and establishes a product with a purity of 99.5 percent from the purchased low-purity bovine serum albumin raw material through anion exchange chromatography, ultrafiltration condensation, and freeze-drying. The invention has few process steps, short process, simple operation, is beneficial to industrialized production and has high product purity.

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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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/765C07K1/18
Inventor 孙永跃何程铖
Owner TIANJIN UNIVERSITY OF TECHNOLOGY
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