DD160582A3 - PREPARATION OF ALBUMIN USING HEAT TRANSFER SYSTEMS - Google Patents

Abstract

The invention relates to a special procedure in the technology of heat fractionation for the production of albumins. The aim of the invention is to reduce the risk of microbial contamination and thus the formation of pyrogenic substances in o.g. To eliminate the method, to shorten the duration of the process, to reduce the heat load of the human plasma and to apply more energetically effective procedures. Areas of application are (industrial) plasma fractionators for human albumin and animal albumins.

Description

220702 ".

Production of albumin using; of heat transfer systems

Field of application of the invention;

The invention relates to a process technology for the isolation of albumin from human and animal serums and plasmas. The method is particularly suitable for all facilities in which the plasma fractionation for the production of infusion albumin is industrymäßig operated.

Characteristic of the known technical solutions

For the isolation of albumin from plasmas and sera on an industrial scale, essentially two methods are used. This is the Cohn method (J.Am.Chem.Soc.68, 459 (1946)) and variants of this method.

It provides in a yield of about 70% albumin having a purity of 95-98% at the end of a 5-stage Fraktionierungsprozessea. Total fractionated separation by the Cohn method requires about 6-8 days of time on normal 8-hour workdays.

Furthermore, all work must be carried out at temperatures below ⁰ ° C.

In contrast, the second route, the heat denaturation of all plasma proteins except the albumin and its separation, represents a decided improvement. (P 2415079) DE, DDR V / P HI 025

220702

With over 90% yield, a product with almost 100% purity is obtained. The time required is based on normal 8-hour days 3 days. The technical effort is significantly reduced in this procedure compared to the Cohn method.

The crucial part of the heat fractionation is to heat the albumin-containing plasma or serum with the addition of alcohol or PEG and substances with stabilizing properties to 60-80 ⁰ C. Under these conditions, the globulins are completely precipitated while the albumin remains in solution. The separation of the globulins is carried out by precoat ion.

The albumin is contained in the filtrate in pure form and is brought either by precipitation reagents or by ultrafiltration to the desired concentration.

Object of the invention

The object of the invention is to improve an important technical sub-step in the heat fractionation process for the production of infusional albumin by avoiding pyrogenic contamination by avoiding bacterial growth during the heating-up time, and by reducing the working time and energy, enabling more effective heat transfer and better control of the denaturing process ,

Explanation of the essence of the invention

In the known methods of heat fractionation, the heating process of the starting material is carried out under constant stirring in double-walled pressure vessels or by heat transfer via heating coils, which are housed in the Rührkesael.

This procedure has the disadvantage that for heating larger plasma volumes of +4 ⁰ C to 68 ⁰ C and 76 ⁰ C are needed up to 5 hours, during this time is in the range between 15-45 ⁰ C, the growth rate of existing micro-organisms especially favors and thus also the education

pyrogenically active bacterial metabolites.

Another disadvantage of this procedure is that the temperature differences between the heating surfaces and the medium must be large, in order to achieve heating in a reasonable time at the relatively small heating surface. This leads to denaturation on the surface and thus to yield losses and quality losses in the infusion albumin.

The energy utilization for heating the plasma or serum is not very effective with these systems, heating coils or double-walled pressure vessels.

These problems in temperature transmission and the consequent disadvantages such as bacterial growth, time expenditure, energy utilization, also occur in the cooling phase.

According to the invention, these disadvantages have been eliminated by the use of heat transfer systems for the isolation of infusion albumin on an industrial scale.

According to the invention, the method is characterized by the following steps:

- Use of pooled plasma with a temperature of +4 ⁰ C - +6 ⁰ C without prior sterile filtration

- Heating the plasma in the presence of polyethylene glycol or alcohol and a stabilizing substance in the flow in a heat exchanger system to 76 ⁰ C and 68 ⁰ C.

- Porting of the heating process in a hot-holding container with stirrer for 30 minutes at 76 ° C or 68 ° C.

- Cooling of the mixture in a 2nd section of the heat exchanger system to ^ j +10 ⁰ C.

Precoat filtration to separate the denatured globulins

- Working up of albumin supernatant to infusion albumin in a known manner

The implementation of the method steps can be carried out according to the following technical variants:

2207UZ

The heating of the plasma or serum is carried out in the flow in a heat exchanger system. Due to the rapid heating time of <N / 8 see. and the even shorter incubation phase between + 15 and + 45 ⁰ C, which only *> / 5 see. claims, a bacterial proliferation is excluded. This allows the use of pooled material without prior sterile filtration. From the heat exchanger system, the heated product passes into a hot-holding container with stirrer, in which it is kept for 30 min. At this temperature. After this time the resulting suspension is cooled in the flow through a heat exchanger system quickly to +10 ⁰ G. The cooling time is about 9 see.

The precipitated globulins are separated by precoat filtration or centrifuges. The albumin can be precipitated from the supernatant with conventional precipitating agents. However, it is more advantageous to carry out purification and concentration of the albumin simultaneously in a diafiltration step.

Embodiment 1

The starting material is donor plasma, which is HB ₃ -Ag negative and has normal transaminase values. Sodium caprylate is added to the starting plasma at a temperature of +4 ⁰ C in a concentration of 0.01 mol, mixed with 3% polyethylene glycol MG 4OOO and adjusted to a pH of 6.5 with 1 η HCl. The mixture is heated in a heat transfer system (17 l / min to 76> ° C) and kept in a holding container with stirring for 30 min. At this temperature. From the holding container, the product is conveyed via the cooling section of the heat exchanger system into a 10 C cooled storage tank with stirrer.

In this, the pH is adjusted to 5.0 with 1 η hydrochloric acid, treated with a filter aid and filtered through a precoat clearly, the filter cake is washed and the clear wash water combined with the filtrate.

2Z07ÜZ

The combined filtrates are filtered through depth filters and added with polyethylene glycol to a content of 22%, wherein the albumin is deposited as a paste. The albumin paste is separated by centrifugation, taken up in distilled water, dialyzed and the desired protein content and osmolality are set. After filtration, the filling into infusion containers and the pasteurization of the albumin takes place for 10 h at 60 ⁰ C with the addition of stabilizers.

Embodiment · 2

Plasma is used as starting plasma as described under 1. This is mixed with sodium caprylate 0.01 mol and 3% methanol. With 1 η hydrochloric acid, the solution is adjusted to a pH of 6.5. In the run, the mixture is heated in a heat exchanger system to 68 ⁰ C and held at this temperature in a holding container with stirring for 30 min. From this, the suspension obtained is cooled to 10 ^° C. in a heat exchanger system. Thereafter, the product is processed as in Example 1.

Claims (1)

(θ Brf indungsans pruch A process for the isolation of albumin from human or animal plasma or blood protein containing fractions in the presence of polyethylene glycol or alcohols and
Stabilizers, characterized in that the plasmas
or heated fractions in heat transfer systems and the heated suspensions are also cooled in heat transfer systems. -9? HV. Ι? Ή