戊二醛化学交联注意事项.docVIP

Glutaraldehyde Glutaraldehyde is the most popular bis-aldehyde homobifunctional crosslinker in use today. However, a glance at glutaraldehyde’s structure is not indicative of the complexity of its possible reaction mechanisms. Reactions with proteins and other amine-containing molecules would be expected to proceed through the formation of Schiff bases. Subsequent reduction with sodium cyanoborohydride or another suitable reductant would yield stable secondary amine linkages (Chapter?3, Sections 4.4 and 5.3). This reaction sequence certainly is possible, but other crosslinking reactions also occur. Glutaraldehyde in aqueous solutions can form polymers containing points of unsaturation due to aldol formation ((Figure 5.25) (Chapter 15, Section 2.1) (Hardy et?al., 1969, 1976;Monsan et al., 1975). Such α,β-unsaturated glutaraldehyde polymers are highlyreactive toward nucleophiles, especially primary amines. Reaction with a protein results in alkylation of available amines, forming stable secondary amine link-ages. These glutaraldehyde modified proteins still may react with other amine-containing molecules either through the Schiff base pathway or through addition at other points of unsaturation (Figure 5.26). The proposed reaction mechanism of conjugation using these polymer conjugates may explain the stability of proteins crosslinked by glutaraldehyde that has not been reduced. Schiff base formation alone would not yield stable crosslinked products without reduction. In addition, a number of other potential reactions of glutaraldehyde in aqueous solution also can contribute to its stable crosslinking ability. These include reactions involving hemiacetal rings sometimes combined with aldol formation products, which can couple to amine groups without the formation of Schiff base linkages (see Chapter?15, Section 2.1, for an in-depth discussion of these reactions). FIGURE 5.24 Two amine-containing molecules can be crosslinked by formaldehyde through formation