RENATURATION OF RECOMBINANT SECRETORY LEUKOCYTE PROTEASE INHIBITOR - ASPECTS OF DISULFIDE BOND FORMATION KINETICS

Therapeutic proteins produced in procaryotic hosts often contain disul fide bonds, which must be fully formed to satisfy United States Food a nd Drug Administration regulations. Native secretory leukocyte proteas e inhibitor (SLPI), a possible emphysema therapeutic agent, contains m any disulfide bonds. However, when SLPI is produced in Escherichia col i by rDNA technology, the disulfide bonds are not formed correctly and must be generated by in vitro renaturation. In this study, the reacti on rate parameters were estimated for SLPI renaturation. The apparent activation energy was approximately 5 kcal/mol suggesting that renatur ation is a diffusion limited process. Apparent reaction rate orders we re not constant, suggesting complex renaturation mechanism(s).