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Method for purification of albumin

A technology for albumin and total protein, applied in the field of purifying albumin, which can solve the problems of aggregate pollution and protein yield reduction.

Pending Publication Date: 2020-01-10
伊沃芙生物制品公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Ethanol precipitation has the disadvantage that some proteins tend to denature in the presence of ethanol, resulting in reduced yields of the protein to be separated and contamination with aggregates that need to be removed before an acceptable therapeutic product can be obtained
[0015] Previous reports did not contribute to understanding the complex relationship between total protein concentration in plasma and the ratio of NaCP to total protein during pasteurization to purify albumin

Method used

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  • Method for purification of albumin
  • Method for purification of albumin
  • Method for purification of albumin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0108] Material

[0109] Starting solution: plasma, partially fractionated plasma (eg plasma depleted of IgG using Cohnfractionation or expanded bed adsorption chromatography).

[0110] NaCP: sodium octanoate, 0.6M stock solution, pH 7.5; Acid: 0.5M-1M stock solution; P300: P300 filter aid; P1000: P1000 filter aid; NaOH: 1M stock solution; Water: WFI; pH meter: Mettler Toledo; Depth filter: Pall K700 depth filter; and fiberglass filter: Pall (10 μm).

[0111] Pasteurization of column D FT+W concentrate in the presence of NaCP

[0112] The column D FT+W concentrated solution was mixed with NaCP, adjusted to pH 7, and then heated at 60°C. The heated solution was cooled to room temperature and then adjusted to pH 5.2 using citric acid.

[0113] The total protein concentration (060716 SPTFF-2) was assumed to be about 1.1 mM. The total protein concentration was estimated by using the molecular weight of albumin (66,500 Da), because by SEC HPLC, the proportion of album...

Embodiment 2

[0127] Pasteurization of column D FT+W concentrate in the presence of NaCP - increasing NaCP concentration and variable bar Sterilization time

[0128] Three different concentrations of NaCP were used such that the final concentration of NaCP in the solution before pH adjustment was 50 mM, 70 mM and 90 mM. The solution was then adjusted to pH 7, followed by pasteurization at 60°C-65°C. Pasteurization at 50 mM and 90 mM NaCP was continued for 4 hours, followed by pH adjustment using citric acid. The 70 mM NaCP condition was aliquoted at 0.5, 1, 2 and 4 hours prior to pH adjustment.

[0129] The total protein concentration (060716SPTFF-2) was assumed to be about 1.1 mM. The NaCP / total protein ratios were 45.5, 63.6 and 81.8 for 50 mM, 70 mM and 90 mM NaCP, respectively. Sample preparation is summarized in Tables 5 and 6. The results of the experiments are expressed as HSA purity based on SEC HPLC analysis (Table 7 and Figure 4 and Figure 5 ). Table 7 shows that inco...

Embodiment 3

[0139] Ratio of NaCP to total protein – three NaCP concentrations to a fixed total protein concentration

[0140] Three different concentrations of NaCP were used such that the final concentration of NaCP in the solution before pH adjustment was 70 mM, 90 mM and 110 mM. The solution was then adjusted to pH 7, followed by pasteurization at 60°C-65°C. Aliquots were taken at 3 hours and 6 hours and the pH was adjusted to 5.2 using citric acid. Prior to centrifugation, the pH adjusted samples were mixed with P-1000 filter aid (0.7% final concentration).

[0141] The total protein concentration (071316SPTFF-2) was assumed to be about 1.8 mM. The NaCP / total protein ratios were 39, 50 and 61 for 70 mM, 90 mM and 110 mM NaCP, respectively. Sample preparation is summarized in Table 8 and Table 9. The results of the experiments are expressed as HSA purity based on SEC HPLC analysis (Table 10 and Figure 7 and Figure 8 ). As shown in Table 9 (see also Figure 7 Results in ), P...

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Abstract

A method for the purification of albumin from plasma is described. The method comprises (a) contacting the plasma with sodium caprylate (NaCP) at an amount dependent on total protein concentration andthe ratio of NaCP to total protein in the plasma, (b) heating the plasma at a near neutral pH range, and (c) separating the albumin from non-albumin-phase. The said method provides for a high yield and purity albumin solution.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of US Provisional Application No. 62 / 472,007, filed March 16, 2017, which is incorporated herein by reference in its entirety. technical field [0003] The present disclosure relates to a new method for purifying albumin from plasma. The method involves contacting the plasma with an amount of sodium caprylate (NaCP) that depends on the concentration of total protein in the plasma and the ratio of NaCP to total protein; subjecting the plasma to a near neutral pH range pasteurization; and separation of albumin from non-albumin phases. The method provides a high yield and purity of the albumin solution. Background technique [0004] Human and animal blood is a source of molecules with therapeutic properties. Many of these molecules are proteins that can be found in plasma or serum. These proteins have been the subject of specific isolation with the aim of purifying and standardizing...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/765A61K38/38C07K1/30C07K1/34C07K1/36
CPCC07K14/765C07K1/30C07K1/34C07K1/36
Inventor J·S·徐D·米勒
Owner 伊沃芙生物制品公司
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