Agarose activation

Supports activated with divinyl sulfone, like those activated with bisoxirane, can be coupled to underivatized carbohydrates.21 The supports activated with divinyl sulfone couple to the hemiacetal hydroxyl group of the carbohydrate by the vinyl group. Accordingly affinity adsorbents prepared by this method will contain glycosyl units. The structure of the reaction product of L-fucose with agarose activated with divinyl sulfone is shown in 5. 

The purification of the pancreatic alpha amylase was effected on an affinity adsorbent prepared from enzymically degraded starch plus agarose activated with bisoxirane.14 The fractions from the affinity column were analyzed for protein components by u.v. absorbance, and for alpha amylase activity by incubating the fractions with starch and measuring the increase in reducing sugars. The results are shown in Fig. 5. 

Another example of the preparation of a special adsorbent is the attachment of L-fucose to agarose activated with divinyl sulfone. This affinity adsorbent was prepared, and used in the purification of two different types of lectin.20 The results on the purification of one lectin on this adsorbent are shown in Fig. 7. 

Fig.l Immobilization of protein ligands on agarose (activation and coupling) by cyanogen bromide. 

Divinylsulphone-cross-linked agarose has been used for the affinity chromatographic purification of an agarase from a culture filtrate oi z Pseudomonas-Viikc bacterium, Agarose activated with divinylsulphone has been treated with D-mannose and the matrix used for the purification of a lectin from Onobrychis viciifolia. 

The derivative obtained when macroporous agarose activated with phloro-glucinol and divinylsulphone reacted with 1,2-diaminoethane is suitable for the attachment of such affinants as methicillin (see p. 468), in which case the product was used in the purification of p-lactamase. ... 

Cyclohepta-amylose couples to macroporous agarose activated with 1,4-bis-(2,3-epoxypropyloxy)butane to give a material that can be used in the purification of a-amylase. ... 

Reaction with an amino-propyl derivative of glass or hornblende with glutaraldehyde cross-linking reaction with titanium(iv) oxide coated with polymerized, diazotized 1,3-diaminobenzene Reaction with agarose activated with 4-benzoquinone Reaction with agarose cyclic imidocarbonate... 

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