Potent inhibition of human immunodeficiency virus type 1 replication by an intracellular anti-Rev single-chain antibody.

Abstract

Medical Sciences. In the article "t(11;22)(q23;q11.2) in acute myeloid leukemia of infant twins fuses MLL with hCDCrel, a cell division cycle gene in the genomic region of deletion in DiGeorge and velocardiofacial syndromes" by Maureen D. Megonigal, Eric F. Rappaport, Douglas H. Jones, Terrence M. Williams, Brian D. Lovett, Kara M. Kelly, Paul H. Lerou, Thomas Moulton, Marcia L. Budarf, and Carolyn A. Felix, which appeared in number 11, May 26, 1998, of Proc. Natl. Acad. Sci. USA (95, pp. 6413-6418), the authors wish to note the following corrections. On page 6414, column 2, line 4, the text should read: "The 24-l ligation reaction mixture contained 0.5 g" not "0.05 g" as printed. Also, on page 6415, column 1, line 56, the text should read: "AmpliTaq DNA polymerase, all four dNTPs (each at 200 M)" not "(each at 250 M)" as printed.Medical Sciences. In the article "Potent inhibition of human immunodeficiency virus type 1 replication by an intracellular anti-Rev single-chain antibody" by Lingxun Duan, Omar Bagasra, Mark A. Laughlin, Joseph W. Oakes, and Roger J. Pomerantz, which appeared in number 11, May 24, 1994, of Proc. Natl. Acad. Sci. USA (91, 5075-5079), the undersigned authors wish to note the following: "The sequence of the heavy chain of the D8 anti-Rev single chain variable fragment (SFv) has been reanalyzed and found to be not what was reported in the article. It is likely that the coding DNA was derived from the fusion partner cell line used to make the original D8 hybridoma, and not from the heavy chain gene expressed by the B cell precursor to this hybridoma. There is a deletion in the framework region 3 (FR3) leading to a framehsift in CDR3 and downstream regions of the heavy chain gene. Individual nucleotide differences make this sequence very close to that described [Thammana, P. (1994)

Intracellular targeting and functional analysis of single-chain Fv fragments in mammalian cells

“…The complete removal of the leader (leaderless), leaving the N-terminal methionine, leads to the localisation of the corresponding protein to the cytoplasm and most likely represents the best choice due to a greater stability of the corresponding protein [5][6][7][8].…”

“…This can be obtained by exploiting the property of dominant and autonomous targeting sequences that can be grafted onto other reporter proteins such as antibodies to confer them a new intracellular localisation. Recombinant antibody domains, in particular single-chain Fv (scFv) 1 fragments, have been successfully expressed inside cells to inhibit the function of several antigens in the cytoplasm [5,6], the nucleus [7,8], and the secretory pathway of mammalian cells [9]. This type of strategy has a great potential for a variety of applications, including gene therapy [10], plant biotechnology [11] and, more recently, functional genomics [4,12,26].…”

Generation and functional characterization of intracellular antibodies interacting with the kinase domain of human EGF receptor

“…This requires alternative stabilizing features of intracellular antibodies to be potent enough to promote correct folding and prevent aggregation under these conditions (Biocca et al, 1995;Proba et al, 1997;Wo¨rn and Plu¨ckthun, 1998;Cattaneo and Biocca, 1999;Zhu et al, 1999). Only in some cases, individual antibody fragments not specifically selected for intracellular expression could be produced as functional cytoplasmic proteins (Biocca et al, 1994;Duan et al, 1994;Mhashilkar et al, 1995;Gargano and Cattaneo, 1997). Therefore, different strategies have been suggested to more reliably generate antibody fragments that are active in the cytosol (Cattaneo and Biocca, 1999;Ohage et al, 1999;Wo¨rn and Plu¨ckthun, 2001).…”

Passive Immunization Against Dental Caries and Periodontal Disease: Development of Recombinant and Human Monoclonal Antibodies

“…I). Improvement of the vector with histidine hexamer tails attached to the ScFv fragment allows for rapid purification by metal ion resin affinity chromatography, This modification has no deleterious effects on in vivo transport and folding or antigen binding (Skerra e cal., 1991 (Duan et al, 1994;Tewari et 6il., 1998) and tumors (Hwu et al, 1993;Chen et cl, 1996) have already been developed.…”