Direct chemical extraction of a recombinant viral coat protein from Escherichia coli at high cell density

The release of protein and DNA from nonrecombinant E. coli JM101 and recombinant E. coli HMS174(DE3) expressing L1 (the major viral coat protein of human papillomavirus type 16) as an inclusion body was demonstrated at high cell density (OD600 = 160). For the nonrecombinant strain, extraction efficiency decreased significantly as cell mass increased, with a high viscosity increase in the postextraction broth. A different dependence on cell concentration was observed for the recombinant strain, with total protein extraction efficiency exceeding 85% for both uninduced and induced cells. Almost complete release of the recombinant L1 protein was achieved at high cell concentration (OD600 = 80 similar to 160) without the use of reducing agent. This greatly extends the concentration range for chemical extraction. (C) 2001 John Wiley & Sons, Inc.