Sevriukov E A, Evseev L P
Vopr Med Khim. 1994 Sep-Oct;40(5):23-5.
Lactate dehydrogenase-C4 (LDH) is a specific cell isoenzyme of spermatogenesis and spermatozoa of man and animals. Due to the lower levels of contaminating proteins, isolation of LDH C4 from spermatozoa had a lot of advantages. LDH C4 was isolated as follows: precipitation with ammonium sulfate, separation of LDH C4 from other LDH isoenzymes using immunosorbents, purification from contaminating proteins of oxamate-Sepharose. Antibodies to LDH1-3 immobilized on tresyl-Sepharose were used as immunosorbents. Elution of LDH C4 from oxamate-Sepharose by means of lithium lactate appeared inadequate therefore LDH C4 was eluted using 0.63 mM sodium pyruvate.
乳酸脱氢酶-C4(LDH)是人和动物精子发生及精子中的一种特异性细胞同工酶。由于污染蛋白水平较低,从精子中分离LDH C4具有诸多优势。LDH C4的分离方法如下:用硫酸铵沉淀,使用免疫吸附剂从其他LDH同工酶中分离出LDH C4,从草氨酸盐-琼脂糖的污染蛋白中进行纯化。固定在三氟甲磺酸-Sepharose上的针对LDH1-3的抗体用作免疫吸附剂。用乳酸锂从草氨酸盐-琼脂糖上洗脱LDH C4似乎不充分,因此使用0.63 mM丙酮酸钠洗脱LDH C4。