PROTEIN-BINDING AFFINITY CHROMATOGRAPHY MEDIA

摘要

Affinity chromatography media are prepared by reacting mono or di-chloro triazine or related dyes with a non-cellulosic solid support matrix containing free hydroxy or amine groups in the presence of an alkali metal hydroxide. In the absence of cyanogen bromide activation, the linkage to the support is entirely via the triazine ring giving high and specific protein-binding capacity. Suitable support matrices include polymers and copolymers of dextrose, dextran and amides and especially agarose. Dye binding levels are much higher than those obtained in the presence of carbonates. Mono-chlorotriazinyl dyes are bound at a pH preferably above 9.5 for about 40-60 hours at room temperature or less at elevated temperatures. Dichlorotriazinyl dyes are bound at pH 8 to 12.5 in a few hours at room temperature. The media may be used for the efficient and often highly specific purification of a wide variety of proteins, including enzymes and blood proteins. Typical ligands used have the structure where R<1> is a sulphonated group derived from anthraquinone, a substituted anthraquinone, an aromatic azo compound or a thiocyanine compound and R<2> is either (a) an organic compound or (b) a chlorine atom. The group R<1> preferably is of formula where R<3>, R<4>, R<5> are sulphonyl groups or hydrogen atoms or alkyl or amino substituted derivatives or where R<6>, R<7> and R<8> are sulphonyl groups or hydrogen atoms and the point of attachment to the triazinyl ring may be any point marked * or a substituted derivative.