Improvement of a Recombinant Anti-Monkey Anti-CD3 Diphtheria Toxin Based Immunotoxin by Yeast Display Affinity Maturation of the scFv
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- 作者：Zhirui Wang ; Geun-Bae Kim ; Jung-Hee Woo ; Yuan Yi Liu ; Askale Mathias ; Scott Stavrou ; David M. Neville ; Jr.
- 刊名：Bioconjugate Chemistry
- 出版年：2007
- 出版时间：May 2007
- 年：2007
- 卷：18
- 期：3
- 页码：947 - 955
- 全文大小：266K
- 年卷期：v.18,no.3(May 2007)
- ISSN：1520-4812
文摘
Recently, a bivalent recombinant anti-human CD3 diphtheria toxin (DT) based immunotoxin derived from thescFv of UCHT1 antibody has been made that shows enhanced bioactivity and is free from the side effects of Fcreceptor interaction. In this case, the diminution of CD3 binding due to the placement of the scFv domain at theC-terminus of the truncated DT in single scFv immunotoxins was compensated by adding an additional scFvdomain. However, this strategy was less successful for constructing an anti-rhesus recombinant immunotoxinderived from the scFv of FN18 antibody due to poor binding of the anti-rhesus bivalent immunotoxin. We reporthere that, by increasing the FN18 scFv affinity through random mutagenesis and selection with a dye-labeledmonkey CD3
recombinant heterodimer, we greatly improved the bioactivity of FN18 derived immunotoxin.The best mutant, C207, contained nine mutations, two of which were located in CDRs that changed the chargefrom negative to positive. Binding affinity of the C207 scFv to the monkey T cell line HSC-F increased 9.8-fold.The potency of the C207 bivalent immunotoxin assayed by inhibition of protein synthesis increased by 238-fold.
recombinant heterodimer, we greatly improved the bioactivity of FN18 derived immunotoxin.The best mutant, C207, contained nine mutations, two of which were located in CDRs that changed the chargefrom negative to positive. Binding affinity of the C207 scFv to the monkey T cell line HSC-F increased 9.8-fold.The potency of the C207 bivalent immunotoxin assayed by inhibition of protein synthesis increased by 238-fold.