Targeting TNF-α with a tetravalent mini-antibody TNF-TeAb

Abstract

Anti-TNF-alpha [anti-(tumour necrosis factor-alpha)] therapy is widely considered to be among the most efficient treatments available for rheumatoid arthritis, psoriatic arthritis and inflammatory bowel disease. In the present study a tetravalent mini-antibody, named 'TNF-TeAb', was constructed by fusing the tetramerization domain of human p53 to the C-terminus of an anti-TNF-scFv [anti-(TNF-alpha-single-chain variable fragment)] via a long and flexible linking peptide derived from human serum albumin. TNF-TeAb was overexpressed as inclusion bodies in the cytoplasm of Escherichia coli, purified to homogeneity by immobilized- metal affinity chromtaography under denaturing conditions and produced in functional form by using an in vitro refolding system. In vitro bioactivity assays suggested that tetramerization of TNF-scFv resulted in an enormous gain in avidity, which endowed TNF-TeAb with a stronger ability to inhibit both receptor binding and cytolytic activity of TNF-alpha. TNF-alpha targeting therapy in rats with collagen-induced arthritis demonstrated that TNF-TeAb provided a much more significant therapeutic effect than did TNF-scFv in suppressing arthritis progression, attenuating inflammation and destruction in joints, and down-regulating pro-inflammatory cytokines and anti-(type II collagen) antibody. The conclusions are therefore (i) that multimerization of the antibody fragment by a self-association peptide is an efficient way to increase its avidity and (ii) that TNF-TeAb has potential applicability for anti-TNF-alpha therapy.

Pharmacological chaperoning: A primer on mechanism and pharmacology

“…In silico docking experiments with CFTR F508del suggest that the VX-809 aromatic ring(s) occupies a hydrophobic pocket at the NBD1/CL4 (cytoplasmic loop) interface [41,42], as well as interact at the NBD1/NBD2 interface and NBD1-CL1/MSD1 interface [42]. Thus, it is likely that correctors that bind to distinct sites on CFTR and additively address multiple structural issues will be required to achieve substantial clinical efficacy, and even with NBD1 corrected and the interfaces restored, potentiators may be needed to achieve full functionality [4,34,43–45]. …”

Inhibition of the development of collagen-induced arthritis in Wistar rats through vagus nerve suspension: a 3-month observation

“…Currently, it is generally accepted that pro-inflammatory cytokines play an important role in the pathogenesis of RA [6][7][8][9]. Therefore, several treatment methods for RA aim to block certain cytokines such as tumor necrosis factor (TNF) and interleukin-1 (IL-1), noted to be essential in the development of RA [10][11][12][13][14][15].…”

The tetravalent anti-DR5 antibody without cross-linking direct induces apoptosis of cancer cells

“…1. The tetravalent antibody was a homotetramer formed association of four scFv fusion products with residues 319-360 of the human p53 [15]. Results of size exclusion HPLC indicated that the purified protein exhibited a major peak (65.5%) with a retention time appropriate for the tetramer and a minor (34.5%) peak with a retention time appropriate for the dimer (Fig.…”

“…The scFv, HSA (human serum albumin) -p53 gene, comprising residues 490-513 of HSA [14] and residues 319-360 of the human p53 [15,16] was assembled by an overlapping PCR with seven synthetic oligonucleotides ( Table 1). The PCR product was cloned into the eukaryotic expression vector pSecTag2A.…”