BACKGROUND: 3B3 is a high-affinity anti-gp120 antibody that neutralizes a wide range of primary and laboratory isolates of HIV-1. The parental antibody was isolated from a combinatorial phage display library constructed from bone marrow RNA of an HIV-infected individual. We have generated a highly active immunotoxin using the 3B3 single-chain Fv (scFv) which can specifically kill lymphocytes infected by HIV-1. MATERIALS AND METHODS: We used recombinant DNA technology to clone the Fv fragment of 3B3 and produce a single-chain Fv (scFv). 3B3 scFv was then fused to a truncated version of Pseudomonas exotoxin A (PE38), giving rise to a recombinant immunotoxin 3B3(Fv)-PE38 that was expressed in E. coli and purified to near homogeneity. RESULTS: 3B3(Fv)-PE38 binds with the same affinity as the parental Fab antibody to the MN strain of gp120. The immunotoxin specifically kills a gp120-expressing transfected cell line and a chronically HIV-infected lymphocytic cell line. The immunotoxin is very stable at 37 degrees C, retaining 80% of its original activity after 24 hr. CONCLUSIONS: Potent immunotoxins such as 3B3(Fv)-PE38 could be utilized in combination with multidrug cocktails that limit viral replication to help reduce viral reservoirs in patients with AIDS.
BACKGROUND: 3B3 is a high-affinity anti-gp120 antibody that neutralizes a wide range of primary and laboratory isolates of HIV-1. The parental antibody was isolated from a combinatorial phage display library constructed from bone marrow RNA of an HIV-infected individual. We have generated a highly active immunotoxin using the 3B3 single-chain Fv (scFv) which can specifically kill lymphocytes infected by HIV-1. MATERIALS AND METHODS: We used recombinant DNA technology to clone the Fv fragment of 3B3 and produce a single-chain Fv (scFv). 3B3 scFv was then fused to a truncated version of Pseudomonas exotoxin A (PE38), giving rise to a recombinant immunotoxin 3B3(Fv)-PE38 that was expressed in E. coli and purified to near homogeneity. RESULTS: 3B3(Fv)-PE38 binds with the same affinity as the parental Fab antibody to the MN strain of gp120. The immunotoxin specifically kills a gp120-expressing transfected cell line and a chronically HIV-infected lymphocytic cell line. The immunotoxin is very stable at 37 degrees C, retaining 80% of its original activity after 24 hr. CONCLUSIONS: Potent immunotoxins such as 3B3(Fv)-PE38 could be utilized in combination with multidrug cocktails that limit viral replication to help reduce viral reservoirs in patients with AIDS.
Authors: W Cavert; D W Notermans; K Staskus; S W Wietgrefe; M Zupancic; K Gebhard; K Henry; Z Q Zhang; R Mills; H McDade; C M Schuwirth; J Goudsmit; S A Danner; A T Haase Journal: Science Date: 1997-05-09 Impact factor: 47.728
Authors: F Barré-Sinoussi; J C Chermann; F Rey; M T Nugeyre; S Chamaret; J Gruest; C Dauguet; C Axler-Blin; F Vézinet-Brun; C Rouzioux; W Rozenbaum; L Montagnier Journal: Science Date: 1983-05-20 Impact factor: 47.728
Authors: Sergey A Trushin; Gary D Bren; Susana Asin; Kevin N Pennington; Carlos V Paya; Andrew D Badley Journal: J Virol Date: 2005-08 Impact factor: 5.103
Authors: Julia A M Sung; Joy Pickeral; Liqin Liu; Sherry A Stanfield-Oakley; Chia-Ying Kao Lam; Carolina Garrido; Justin Pollara; Celia LaBranche; Mattia Bonsignori; M Anthony Moody; Yinhua Yang; Robert Parks; Nancie Archin; Brigitte Allard; Jennifer Kirchherr; JoAnn D Kuruc; Cynthia L Gay; Myron S Cohen; Christina Ochsenbauer; Kelly Soderberg; Hua-Xin Liao; David Montefiori; Paul Moore; Syd Johnson; Scott Koenig; Barton F Haynes; Jeffrey L Nordstrom; David M Margolis; Guido Ferrari Journal: J Clin Invest Date: 2015-09-28 Impact factor: 14.808