Figure 1.
Preparation and characterization of L19mTNFα fusion protein and its control fusion protein TN11mTNFα (A) Scheme of the homotrimeric form of the fusion protein scFv-mTNFα in which the mTNFα monomers are held together through hydrophobic interactions. (B) Scheme of the construct for L19mTNFα and TN11mTNFα inserted in the mammalian expression vector pCDNA3.1, under the control of the cytomegalovirus (CMV) promoter. (C) SDS-PAGE analysis of the purified fusion proteins L19mTNFα (lane 1) and TN11mTNFα (lane 2). As expected, a major band corresponding to the monomer of about 45 kDa was detected. Numbers on the right are the apparent molecular masses of the standard, in kilodaltons. (D) Gel filtration profile of the purified L19mTNFα and TN11mTNFα (Superdex 200). The retention volume (milliliter) of the peaks corresponds to an apparent molecular mass of about 140 kDa, consistent with the homotrimeric format of the 2 fusion proteins.