Luley C, Prellwitz W, Oster O, Kloer H U
Anal Biochem. 1987 May 15;163(1):182-7. doi: 10.1016/0003-2697(87)90110-2.
Isoelectric focusing (IEF) of apolipoproteins is an important procedure for the isolation of apolipoprotein isoforms. In this paper we describe the use of agarose/urea as a focusing medium for IEF of apolipoproteins. This technique, which offers several advantages over the traditional polyacrylamide gel focusing, yields a further microheterogeneity of the known isoforms of apolipoproteins A-I and E. In addition, by subsequent immunoprecipitation, both specificity and sensitivity are enhanced. Using a monospecific antiserum against Apo A-I, a new minor isoform was detected at the basic end of the Apo A-I spectrum (pI 6.02) which has not yet been observed in normal plasma samples. Another advantage of this technique in combination with immunoprecipitation is the specific detection of Apo E phenotypes which is essential for the diagnosis of hyperlipoproteinemia type III.
载脂蛋白的等电聚焦（IEF）是分离载脂蛋白异构体的重要方法。在本文中，我们描述了使用琼脂糖/尿素作为载脂蛋白IEF的聚焦介质。该技术比传统的聚丙烯酰胺凝胶聚焦具有多个优势，可产生载脂蛋白A-I和E已知异构体的进一步微异质性。此外，通过后续的免疫沉淀，特异性和灵敏度均得到提高。使用针对Apo A-I的单特异性抗血清，在Apo A-I谱的碱性端（pI 6.02）检测到一种新的次要异构体，这在正常血浆样本中尚未观察到。该技术与免疫沉淀相结合的另一个优势是可特异性检测Apo E表型，这对于III型高脂蛋白血症的诊断至关重要。
