Leah J M, Billett E E, Palmer T
Department of Life Sciences, Trent Polytechnic, Nottingham, United Kingdom.
Anal Biochem. 1988 May 1;170(2):495-501. doi: 10.1016/0003-2697(88)90664-1.
Ornithine aminotransferase was purified by conventional biochemical methods from rat kidney, rat liver, and human liver. Affinity-purified antibodies raised to the rat kidney enzyme were used to produce an immunoadsorbent enabling a one-step purification of ornithine aminotransferase to be made from crude human liver extracts. The harsh chemical conditions often required to desorb immunoadsorbents were avoided by isolating antibodies with low functional affinity and employing an electrophoretic desorption method which allowed the enzyme activity to be retained. The close structural similarity between human and rat ornithine aminotransferase was demonstrated by immunodiffusion reactions. It was therefore possible to purify the enzyme from human liver using immobilized antibodies raised against rat kidney ornithine aminotransferase. Furthermore, desorption was more readily achieved due to the lower affinity for the human enzyme.
采用常规生化方法从大鼠肾脏、大鼠肝脏和人类肝脏中纯化鸟氨酸转氨酶。用针对大鼠肾脏酶产生的亲和纯化抗体来制备免疫吸附剂，从而能够从人肝脏粗提物中一步纯化鸟氨酸转氨酶。通过分离低功能亲和力的抗体并采用能保留酶活性的电泳解吸方法，避免了通常解吸免疫吸附剂所需的苛刻化学条件。免疫扩散反应证明了人和大鼠鸟氨酸转氨酶之间存在密切的结构相似性。因此，有可能使用针对大鼠肾脏鸟氨酸转氨酶产生的固定化抗体从人肝脏中纯化该酶。此外，由于对人酶的亲和力较低，更容易实现解吸。
