Use of rabbit Fab'-peroxidase conjugates prepared by the maleimide method for detecting plant viruses by ELISA

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.