Degl'Innocenti D, Berti A, Stefani M, Liguri G, Ramponi G
Department of Biochemical Sciences, University of Florence, Italy.
Biotechnol Appl Biochem. 1990 Aug;12(4):450-9.
Specific anti-human erythrocyte acylphosphatase antibodies were raised in rabbits, purified by affinity chromatography, and used to develop an enzyme purification procedure based on an immunoaffinity chromatography step. This procedure permitted the rapid purification of the enzyme, with a high final yield and with a specific activity very similar to that found for the enzyme purified by the standard procedure. The noncompetitive enzyme-linked immunoadsorbent assay developed with the affinity-purified antibodies was very specific and sensitive in that a positive reaction could be detected in the presence of antigen amounts of as little as 0.01 ng/ml. By this assay the enzyme content was determined in normal cells, tissues, and organs as well as in blood samples from hemopathy-affected patients. This test could possibly have clinical applications.
在兔体内产生了特异性抗人红细胞酰基磷酸酶抗体，通过亲和层析进行纯化，并用于开发基于免疫亲和层析步骤的酶纯化程序。该程序能够快速纯化该酶，最终产率高，比活性与通过标准程序纯化的酶非常相似。用亲和纯化抗体开发的非竞争性酶联免疫吸附测定法非常特异且灵敏，在抗原量低至0.01 ng/ml时就能检测到阳性反应。通过该测定法，测定了正常细胞、组织和器官以及血液病患者血样中的酶含量。该检测可能具有临床应用价值。
