DE20109829U1 - Filtration extraction system - Google Patents

Description

Filtration extraction system

The invention relates to a filtration-extraction system in 96-well or 384-well format for biotechnological work, preferably in protein and nucleic acid technology, consisting of several individual parts that are optionally connected to one another. In detail, it consists of a collecting plate with a knobbed lid, a positioning frame, an adapter plate, a filtration plate that contains a matrix with bound macromolecules and is closed with a foil lid.

Extraction systems in the form of automatic extraction machines, in which the gel samples are extracted in flow reaction chambers, are known per se. However, they have the disadvantage that small pieces of gel can be lost. Furthermore, they are characterized by low throughput speeds (approx. 95 samples per day) and their purchase price is high.

The method currently most commonly used for extracting proteins involves placing a piece of gel in a single reaction vessel, adding the solutions, centrifuging and removing the supernatant. This method also has numerous disadvantages, as it is labor-intensive, slow (approx. 30 samples per day) and small pieces of gel can also be lost due to careless work.

The invention was therefore based on the object of providing an extraction system that is safe and cost-effective and ensures a high daily throughput for biomolecules to be extracted.

The filtration-extraction system according to the invention in 96-well or 384-well format, preferably for the cleavage of proteins and simultaneous extraction of peptides, is composed of various components and consists of

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essentially consists of a collecting plate, possibly with a knobbed lid, a positioning frame, an adapter plate, a filtration plate containing a matrix with bound macromolecules and a foil lid for the filtration plate.

The cross-section of such a system is shown in Fig. 1:

1 collecting plate

2 Filtration plate

3 knobbed lids for collecting plate

4 Positioning frames

5 Adapter plate

6 foil lids for filtration plate

7 Matrix, preferably with proteins (bound or contained)

The core of the system is the so-called filtration plate, which is a known 96-well or 384-well microtiter plate with a filtration base, whereby the pores of the filtration base are in the με-βββα range. The matrix, to which the macromolecules to be split, preferably proteins, can be bound or which contains the macromolecules (e.g. protein gel piece), is placed in the wells of the filtration plate to carry out the process.

The filtration and extraction process is carried out by centrifugation with special washing and extraction solutions. One difficulty with similar systems is that the matrix becomes electrostatically charged, particularly when drying, and can jump out of the filtration plate when the lid is opened. This is prevented by the matrix being immobilized on the filtration base by high-speed centrifugation. The process is as follows:

A solution is pipetted into the wells of the filtration plate, the filtration bottom of which is tight without the application of pressure. After an incubation period, the solution is

Centrifugation or suction filtering, whereby the matrix remains in the well of the filtration plate while the filtrate is collected in the collecting plate below. In the case of washing steps, the filtrate is discarded; in extraction steps, the filtrates are collected and usually combined.

For example, to split proteins in gel pieces, the following steps are carried out: washing, decolorization, dehydration, rehydration, splitting, extractions.
The extraction solutions dissolve the peptides (protein breakdown products) from the matrix. The extraction solutions are collected in the collection plate and the volume of the extracts is reduced in an evaporator.

With this device, immobilized proteins can be cleaved and the resulting peptides extracted and subjected to identification (e.g. mass spectroscopy).

In addition to the components shown in Fig. 1, the filtration-extraction system also includes all washing and extraction solutions required to carry out the process.

Claims (2)

1. Filtration-extraction system for splitting biomacromolecules, in particular for splitting proteins, characterized by at least one collecting plate ( 1 ), a filtration plate ( 2 ), optionally a knobbed lid ( 3 ), optionally a positioning frame ( 4 ), optionally an adapter plate ( 5 ), optionally a film lid for the filtration plate ( 6 ) and a matrix ( 7 ) with biomacromolecules. 2. Filtration-extraction system according to claim 1, characterized in that it comprises all washing and extraction solutions for the cleavage and extraction of the biomacromolecules.