Specificity in the combination of Fd fragments with L chains to form hapten-binding sites.
In the work reported here we have shown that light chains and Fd fragments can be separated completely in propionic acid and then recombined to form Fab fragments with antibody activity. This experiment indicates that in the recombination a correct alignment of the Fd fragments and the L chains occurs to give a competent antibody site, just as occurs with the recombination of separated heavy and light chains of the antibody; thus the Fc fragment is not required for correct alignment. Fd fragments of antibody alone show very low binding activity toward the specific hapten. As is the case for the combination of heavy and light chains, the combination of Fd fragments and light chains also requires that both components come from antibody from the same rabbit in order to give binding sites. When they are derived from different rabbits producing antibody against the same antigen, they still give Fab fragments as shown by immunoelectrophoresis but do not have competent binding sites. An important observation is that the subunits of the papain digest fractions, Fab(I) and Fab(II), have the capacity to cross-combine to form active Fab fragments with competent binding sites. Fd(I) from Fab(I) combines with L(II) chains from Fab(II) to give the composite (Fd(I)-L(II)) with good binding activity. Likewise, the composite (Fd(II)-L(I)) has good binding activity. The composites from the two types of antibody molecules yielding different Fab fragments have antibody activity although heretofore these molecules have appeared to be different on the bases of chromatography and amino acid analysis. There is also a preferential combination of the Fd fragments to combine with the correct L fragments to give binding sites since this combination takes preference over the combination of Fd fragments of antibody with light chains of normal globulin (or of light chains of antibody with Fd fragments of normal globulin).
D000818:Animals; D000906:Antibodies; D002845:Chromatography; D006241:Haptens; D007122:Immunoelectrophoresis; D066298:In Vitro Techniques; D010455:Peptides; D011817:Rabbits; D005719:gamma-Globulins
D000906:Antibodies; D006241:Haptens; D010455:Peptides; D005719:gamma-Globulins