Weiss E, Van Regenmortel M H
Institut de Biologie Moléculaire et Cellulaire, Laboratoire d'Immunochimie, Strasbourg, France.
J Virol Methods. 1989 Apr-May;24(1-2):11-25. doi: 10.1016/0166-0934(89)90003-7.
In contrast to antibodies conjugated to enzyme with glutaraldehyde or by the periodate method, monomeric Fab' fragments conjugated to enzyme by means of a maleimide compound are not adversely affected by the conjugation procedure. We used such Fab'-enzyme conjugates prepared with antibody to tobacco mosaic virus (TMV), to TMV coat protein and to rabbit IgG for the detection of different tobamoviruses by direct and indirect double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Compared to conjugates prepared by other methods, the sensitivity of TMV detection with Fab'-enzyme conjugates by direct DAS-ELISA was markedly increased. However, because of their monomeric nature, these FAb'-enzyme conjugates did not cross-react with serologically related tobamoviruses. Anti-globulin Fab'-enzyme conjugate was found to be the most efficient anti-globulin conjugate for detecting TMV by indirect DAS-ELISA. Because of their high sensitivity and serotype specificity, FAb'-enzyme conjugates are useful for detecting low amounts of contaminating viruses present in crude viral preparations.
与通过戊二醛或高碘酸盐法与酶偶联的抗体不同，通过马来酰亚胺化合物与酶偶联的单体Fab'片段不会受到偶联过程的不利影响。我们使用了用抗烟草花叶病毒（TMV）、TMV外壳蛋白和兔IgG的抗体制备的此类Fab'-酶偶联物，通过直接和间接双抗体夹心酶联免疫吸附测定（DAS-ELISA）来检测不同的烟草花叶病毒。与通过其他方法制备的偶联物相比，通过直接DAS-ELISA用Fab'-酶偶联物检测TMV的灵敏度显著提高。然而，由于其单体性质，这些Fab'-酶偶联物与血清学相关的烟草花叶病毒不会发生交叉反应。发现抗球蛋白Fab'-酶偶联物是通过间接DAS-ELISA检测TMV最有效的抗球蛋白偶联物。由于其高灵敏度和血清型特异性，Fab'-酶偶联物可用于检测粗病毒制剂中存在的少量污染病毒。
