Persoon N L, Sips H J, Hülsmann W C, Jansen H
Biochim Biophys Acta. 1986 Feb 12;875(2):286-92. doi: 10.1016/0005-2760(86)90178-5.
To obtain monoclonal antibodies against rat salt-resistant liver lipase, mice were immunized with enzyme purified from heparin-containing rat liver perfusates. Hybridomas were screened for antibody production by means of an enzyme-linked immunosorbent assay (ELISA) and an immunoprecipitation assay. Five hybridoma cell lines secreting antibodies against rat liver lipase indicated as A, B, C, D and E, have been obtained. All antibodies possess gamma one (gamma 1) heavy chains and kappa (kappa) light chains. The antibodies precipitate salt-resistant lipase from rat post-heparin plasma, are positive in ELISA, inhibit liver lipase activity and bind monospecifically with the enzyme as shown by immunoblotting. The monoclonal antibodies showed no significant reactivity with human liver lipase. The salt-resistant lipases of rat adrenals and ovaries are also precipitated by the monoclonal antibodies directed against the liver enzyme. Therefore, the heparin-releasable lipases of the liver, adrenals and ovaries possess identical epitopes.
为获得抗大鼠耐盐肝脂肪酶的单克隆抗体，用从含肝素的大鼠肝脏灌注液中纯化的酶免疫小鼠。通过酶联免疫吸附测定（ELISA）和免疫沉淀测定筛选杂交瘤的抗体产生情况。已获得五个分泌抗大鼠肝脂肪酶抗体的杂交瘤细胞系，分别标记为A、B、C、D和E。所有抗体均具有γ1重链和κ轻链。这些抗体可从大鼠肝素后血浆中沉淀出耐盐脂肪酶，在ELISA中呈阳性，抑制肝脂肪酶活性，并如免疫印迹所示与该酶特异性结合。这些单克隆抗体与人肝脂肪酶无明显反应性。针对肝脏酶的单克隆抗体也能沉淀大鼠肾上腺和卵巢的耐盐脂肪酶。因此，肝脏、肾上腺和卵巢中可被肝素释放的脂肪酶具有相同的表位。
