Recovery of Antibody Activity from Inactive Hybrids

Globulin molecules can be reconstituted from th e separated heavy (H) and light (L) chains of specifically purified antihapten antibody (1, 2). When the chains are derived from the same rabbit, antibody activity is recovered (1, 3, 4). Hybrid molecules of chains from different rabbits have been found to show only questionable activity (5, 6). Since the chains in hybrids are not linked by covalent bonds, we have now investigated their stability toward dissociation. We found that the hybrids in a mixture consisting of one hybrid having H chains from the anti-p-azobenzoate (anti-Xp) x antibody of one rabbit and L chains from the anti-p-azobenzenearsonate (anti-Rp) 1 antibody of a second rabbit and a second hybrid of the opposite combination are so stable at pH 8 and at 5°C that even after 2 wk only low residual binding activity toward either specific hapten was shown. Exposure of the mixture to 1 ~ propionlc acid followed by buffer resulted in a major recovery of binding activity toward haptens specific for anti-Xp and for anti-Rp. These results show that no exchange occurred between the hybrids at pH 8, but that exchange did occur on exposure to propionic acid and with a preferential recombination of the chains to give effective antibody sites.