Catalyst – solid sorbent – or support therefor: product or process – Solid sorbent – Organic
Patent
1983-12-23
1985-07-30
Konopka, P. E.
Catalyst, solid sorbent, or support therefor: product or process
Solid sorbent
Organic
210502, 210656, 21050027, 435176, 435177, 435276, 502 7, B01J 2024, B01D 1508, G01N 3354
Patent
active
045322325
DESCRIPTION:
BRIEF SUMMARY
This invention relates to a separation agent consisting of a water insoluble solid substance containing a covalently bound lectin.
Lectins, especially Concanavalin A, have earlier been bound to solid substances such as agarose. The resulting material has been used for the separation of various carbohydrates, carbohydrate-related substances, organelles and even whole cells (Concanavalin A as Tool, H. Bittiger and H. P. Schnebli, John Wiley and Sons, New York, 1976).
The polymers which have been used as a basic material in these separation agents, for example agarose and polymethacrylic acid, have the drawback of leading to bad flow characteristics due to poor rigidity of the polymer and bad separation characteristics.
According to the present invention there has now been provided a separation agent of the above-mentioned type, which has superior flow and separation characteristics and which is well suited for use in connection with for example high performance liquid chromatography (HPLC). The separation agent according to the invention is characterized in that the solid substance structurally is composed of a water insoluble silica material or silicate material as a basic structure and via a linkage to silicon atoms in said silica or silicate material, a covalently bound lectin.
The expression "silica material" is herein intended to mean silicic acid in more or less dehydrated form, including silica. The basic material may be non-porous or porous silica or glass particles, which are known per se as carrier material in several applications, for example in biochemical determination methods, but can also consist of the inner wall of a tube or be a disk. The lectin is covalently bound to an alkylene chain which contains 1-22, preferably 2-18, carbon atoms and optionally being interrupted by one or more oxygen, sulphur or nitrogen atoms and/or being substituted with one or more hydroxyl groups. Except for the case of amide and ester groups, preferably at most 1 atom of a kind other than carbon and hydrogen is bound to one and the same carbon atom in the chain. The alkylene chain is in turn bound, optionally via an intermediate silicon atom, preferably to an oxygen atom belonging to a hydroxyl group, said hydroxyl group being bound to a silicon atom in the basic material for the present separation agent. The alkylene chain may also be optionally bound directly to a silicon atom in said basic material.
The introduction of the bridge-forming alkylene chain structure and the lectin in the silica or silicate material is preferably performed in two steps. A substituent which corresponds to the alkylene chain and which contains a reactive structure such as for example an epoxy or carboxyl group is introduced in the first step. In the second step the lectin is added which can react with the functional structure on the silica. In some cases a condensation-promoting agent such as a carbodiimide may also be added.
The invention will be illustrated by means of examples in the following text.
EXAMPLE 1
Preparation of Silica Comprising Epoxy Groups
To 20 g porous silica (LiChrosorb Si 1000, 10 .mu.m, E Merck, Darmstadt, Federal Republic of Germany), dried at 200.degree. C. over night and then suspended in 500 ml Na-dried toluene was added 200 microliter triethylamine (potassium hydroxide-dried) and 10 ml .gamma.-glycidoxy-propyltrimethoxysilane. The mixture was reflux boiled under nitrogen for 16 hr. in a flask fitted with a teflon stirrer. Thereafter the substituted silica was filtered off and washed on a glass filter funnel with toluene, acetone and diethyl ether and finally dried at reduced pressure. A sample of the obtained product was analyzed with respect to epoxy groups (reaction with 3 M sodium thiosulphate for 1 hr. at pH 7 and determination of librated hydroxyl ions by addition of acid (pH-stat; 0.10 M hydrochloric acid)). Materials based on LiChrosorb Si 1000 contained about 50 .mu.mole epoxy groups/g.
EXAMPLE 2
Preparation of Silica Containing Diol Groups
3 g silica containing epoxy groups (see exampl
REFERENCES:
patent: 4406792 (1983-09-01), Glad et al.
patent: 4446275 (1984-05-01), Filka et al.
Borchert Axel
Larsson Per-Olof
Mosbach Klaus
Konopka P. E.
Pernovo AB
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