Purification of Leucyl-tRNA synthetase and preparation of monoclonal antibodies
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Leucyl tRNA synthetase (LeuRS) has been purified several hundred-fold from Chinese hamster liver. The purification scheme was as follows: differential centrifugation, Alumina C gamma adsorption, DE-52 anion exchange column chromatography, and finally IGGF cation exchange column chromatography. The partially purified LueRS fraction obtained above was used as antigen in the production of monoclonal antibodies. Two immunization procedures were used. The in vivo procedure involved biweekly intraperitoneal injections of antigen in complete Freund's adjuvant. The in vitro procedure was done in 25 cm² culture flasks and the medium contained a synthetic peptidyl adjuvant. Using either procedure, the relative purity of the LeuRS fraction was important in the efficiency of specific anti- LeuRS clones produced.