Targeting TNF-alpha with a tetravalent mini-antibody TNF-TeAb.
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.
Liu
Mengyuan
M
Faculty of Life Science, Hubei University, 430062 Wuhan, People's Republic of China.
Wang
Xiangbin
X
Yin
Changcheng
C
Zhang
Zhong
Z
Lin
Qing
Q
Zhen
Yongsu
Y
Huang
Hualiang
H
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism / Q000627:therapeutic use; D001168:Arthritis / Q000139:chemically induced / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D001483:Base Sequence; D002460:Cell Line; D002850:Chromatography, Gel; D003094:Collagen / Q000494:pharmacology; D015870:Gene Expression; D007167:Immunotherapy; D053583:Interleukin-1beta / Q000097:blood; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats; D014409:Tumor Necrosis Factor-alpha / Q000097:blood / Q000276:immunology*
D000906:Antibodies; D053583:Interleukin-1beta; D014409:Tumor Necrosis Factor-alpha; D003094:Collagen