Liquid purification or separation – Magnetic – With additional separator
Patent
1986-07-24
1989-08-08
Jones, W. Gary
Liquid purification or separation
Magnetic
With additional separator
210222, 2105021, 435178, 435180, B03C 130
Patent
active
048550450
DESCRIPTION:
BRIEF SUMMARY
The invention relates to a method for the separation of organic substances, more especially cells, organelles and proteins, from a suspension or solution flowing through a reaction space by attaching the organic substances to attachment bodies inside the reaction space in the form of pearls or the like, whose surfaces may be provided with an absorption layer which acts specifically on the respective organic substance.
For the separation of biological cells or organelles, it is known to use pearl-shaped attachment bodies, whose surfaces mainly bind specific cells or organelles. This can be achieved in that special antibodies, haptenes or chemical substrates are covalently attached to the surface of the attachment bodies which, in turn, exclusively or mainly bind special cells or organelles. Once the attachment of the cells or organelles has been effected, the substances not bound to the surface of the attachment bodies can be washed away; subsequently, the substances bound by the attachment bodies can be detached from the attachment bodies with the aid of appropriate methods.
In a known method of the kind mentioned at the beginning, the attachment bodies are contained in a dense package within a so-called separating column, through which the suspension containing the organic substances flows. This arrangement has the disadvantage that, on account of the attachment bodies being packed densely, the separating column offers a high degree of flow resistance so that the suspension flows through the separating column at a low flow velocity. This results in a correspondingly long sojourn time of at least half an hour and up to 2 or 3 hours. This long sojourn time is disadvantageous not only because it impairs the economy of the method but also because this increases the probability of undesirable substances being attached to the attachment bodies in a non-specific bond in such a way that they are not removed in the subsequent normal washing process step. The product finally separated from the absorption bodies therefore contains not only the desired special cells or organelles. Another disadvantage of the known method lies in the fact that cells or organelles and proteins, including the undesirable non-specific cells, organelles or proteins, may be enclosed in the small spaces between the attachment bodies which are in abutting contact with one another. On account of these disadvantages, the use of this method has been confined to preparatory quantities of organic substances to be separated. An enlargement of the plant for cell or organelle suspensions of more than 50 or 100 ml presented considerable difficulties.
It is therefore the object of the invention to reduce the throughput time in a process of the kind mentioned at the beginning without impairing the separating effectiveness.
This problem is solved in that the magnetic, preferably ferromagnetic attachment bodies are kept at a distance from one another in an open arrangement in the reaction space by magnetic field gradients in a ratio of the total volume of the attachment bodies to the reaction space volume of between 0.01 and 0.5, preferably between 0.02 to 0.4, and ideally between 0.03 and 0.2. Due to the distribution of the attachment bodies over the reaction space at a distance from one another in the stated volume ratios, there comes about a low flow resistance which allows a correspondingly high flow velocity of the suspension (i.e. the suspension containing the organic substances). It is virtually impossible for organic substances getting stuck between adjacent attachment bodies. The short throughput time allows the process to be extended beyond a laboratory operation. On account of the shortened sojourn time, the risk of undesirable cells or organelles being bound non-specifically to the attachment bodies is reduced. Due to the relatively high flow velocity, the non-specific cells or organelles are as a rule taken along by the flow so that the subsequent washing process step for removing these substances from the attachment bodies can be dispensed
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