Stults N L, Asta L M, Lee Y C
Department of Biology, Johns Hopkins University, Baltimore, Maryland 21218.
Anal Biochem. 1989 Jul;180(1):114-9. doi: 10.1016/0003-2697(89)90097-3.
Mild periodate oxidation of certain commercially available crosslinked agarose beads (Sepharose CL-4B and CL-6B) results in the generation of aldehydo groups which were useful for immobilization of amino compounds by reductive amination using pyridine borane. Consumption of periodate ion and production of formaldehyde were only observed with crosslinked Sepharose preparations and were correlated with a binding capacity much greater than that of uncross-linked gels when subjected to the reductive amination reaction. Up to 50 mg (approximately 0.73 mumol) of bovine serum albumin and 30 mumol of glycylglycine were coupled per gram of moist oxidized Sepharose CL-6B. The immobilization reaction was shown to proceed at neutral pH requiring about 12 h for completion and to be relatively insensitive to temperature and pyridine borane concentration. The oxidized gel was shown to be stable for at least 2 months upon storage in 0.1 M acetic acid. This method has proven to be useful for the preparation of a variety of affinity matrices and immobilized enzymes.
对某些市售交联琼脂糖珠（琼脂糖CL - 4B和CL - 6B）进行轻度高碘酸盐氧化，会产生醛基，这些醛基可通过使用吡啶硼烷的还原胺化反应来固定氨基化合物。仅在交联琼脂糖制剂中观察到高碘酸根离子的消耗和甲醛的产生，并且在进行还原胺化反应时，其结合能力比未交联的凝胶大得多。每克湿的氧化琼脂糖CL - 6B可偶联高达50毫克（约0.73微摩尔）的牛血清白蛋白和30微摩尔的甘氨酰甘氨酸。固定化反应显示在中性pH下进行，约需12小时完成，并且对温度和吡啶硼烷浓度相对不敏感。氧化凝胶在0.1 M乙酸中储存时显示至少2个月稳定。该方法已被证明可用于制备各种亲和基质和固定化酶。
