Pereyra Ali, Agundis Concepcion, Barrera Baltazar, Alpuche Juan, Sierra Claudia, Zenteno Roberto, Zenteno Edgar, Vazquez Lorena
Laboratorio de Inmunologia, Departamento de Bioquimica, UNAM, Mexico.
Prep Biochem Biotechnol. 2009;39(3):308-22. doi: 10.1080/10826060902953384.
We determined the cross-reactivity of a monoclonal antibody against the Macrobrachium rosenbergii lectin with proteins in the hemolymph from Procambarus clarkii (Pc), Procambarus americanus (Pa), Litopenaeus setiferus (Ls), and Pseudothelphusa americana (Psa). Crustaceans' hemolymph agglutinated erythrocytes from rat, mouse, guinea pig, and rabbit. Decapods' hemolymph hemagglutinating activity was inhibited by N-acetylated carbohydrates as well as by antibodies. Western blot assays indicated that the antibodies recognized two main proteins of 97.5 and 80.9 kDa in all hemolymphs studied; moreover, ELISA assays indicated that, in PSa, 7.2% of total proteins showed crossreactivity with antibodies in Pa, Pc, and Lc hemolymphs represented 4.2, 3.1%, and 2.5%, respectively. Our results suggested that antibodies recognize the lectin active site in the crustacean species tested; we propose the use of antibodies as an immunological marker for lectin identification and quantification among crustaceans.
我们测定了一种针对罗氏沼虾凝集素的单克隆抗体与克氏原螯虾(Pc)、美洲原螯虾(Pa)、凡纳滨对虾(Ls)和美洲伪溪蟹(Psa)血淋巴中蛋白质的交叉反应性。甲壳类动物的血淋巴能凝集大鼠、小鼠、豚鼠和兔子的红细胞。十足目动物的血淋巴凝血活性受到N - 乙酰化碳水化合物以及抗体的抑制。蛋白质印迹分析表明,这些抗体在所研究的所有血淋巴中识别出两种主要蛋白质,分子量分别为97.5 kDa和80.9 kDa;此外,酶联免疫吸附测定表明,在Psa中,总蛋白的7.2%与Pa、Pc和Lc血淋巴中的抗体呈现交叉反应,在Pa、Pc和Lc中分别占4.2%、3.1%和2.5%。我们的结果表明,抗体识别所测试甲壳类物种中的凝集素活性位点;我们建议将抗体用作甲壳类动物中凝集素鉴定和定量的免疫标记物。