Figure 4

Expression-secretion analysis of 16 scFv’s in vectors pLTEX-2 to pLTEX-5. (A) Western blot analysis of scFv’s in culture supernatant. Detection of scFv’s with Protein L-HRP (1:5000). (B) SDS-PAGE analysis of Ni-NTA purified scFv’s from 100 ml cultures. Proteins are separated on 4-12% Bis-Tris gels (Invitrogen). (C) Protein yield from 100 ml culture volumes grown for 72 h of IMAC-purified scFv’s determined in duplicate by densitometrical analysis on SDS-PAGE. Black circles, squares, triangles and stars represent scFv-1 to −4, respectively.