The sub-nanomolar binding of DNA-RNA hybrids by the single-chain Fv fragment of antibody S9.6

被引：122

作者：

Phillips, Damilola D. ^{[1
]}

Garboczi, David N. ^{[2
]}

Singh, Kavita ^{[2
]}

Hu, Zonglin ^{[1
]}

Leppla, Stephen H. ^{[1
]}

Leysath, Clinton E. ^{[1
]}

机构：

[1] NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA

[2] NIAID, Struct Biol Sect, Res Technol Branch, NIH, Rockville, MD 20852 USA

来源：

JOURNAL OF MOLECULAR RECOGNITION | 2013年 / 26卷 / 08期

关键词：

Monoclonal antibody; scFv; antibody sequence; DNA-RNA hybrids; SOLUTION HYBRIDIZATION ASSAY; MONOCLONAL-ANTIBODY; RIBOSOMAL-RNA; HYPERVARIABLE REGIONS; IMMUNODETECTION; BACTERIA;

D O I：

10.1002/jmr.2284

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

070307 [化学生物学]; 071010 [生物化学与分子生物学];

摘要：

The monoclonal antibody S9.6 binds DNA-RNA hybrids with high affinity, making it useful in research and diagnostic applications, such as in microarrays and in the detection of R-loops. A single-chain variable fragment (scFv) of S9.6 was produced, and its affinities for various synthetic nucleic acid hybrids were measured by surface plasmon resonance (SPR). S9.6 exhibits dissociation constants of approximately 0.6nM for DNA-RNA and, surprisingly, 2.7nM for RNA-RNA hybrids that are AU-rich. The affinity of the S9.6 scFv did not appear to be strongly influenced by various buffer conditions or by ionic strength below 500mM NaCl. The smallest epitope that was strongly bound by the S9.6 scFv contained six base pairs of DNA-RNA hybrid. Published 2013. This article is a U.S. Government work and is in the public domain in the USA.

引用

页码：376 / 381

页数：6

共 27 条

[1]

CHARACTERIZATION OF MONOCLONAL-ANTIBODY TO DNA.RNA AND ITS APPLICATION TO IMMUNODETECTION OF HYBRIDS [J].

BOGUSLAWSKI, SJ ;

SMITH, DE ;

MICHALAK, MA ;

MICKELSON, KE ;

YEHLE, CO ;

PATTERSON, WL ;

CARRICO, RJ .

JOURNAL OF IMMUNOLOGICAL METHODS, 1986, 89 (01) :123-130

[2]

MONOCLONAL-ANTIBODY SOLUTION HYBRIDIZATION ASSAY FOR DETECTION OF MOUSE HEPATITIS-VIRUS INFECTION [J].

CASEBOLT, DB ;

STEPHENSEN, CB .

JOURNAL OF CLINICAL MICROBIOLOGY, 1992, 30 (03) :608-612

[3]

CANONICAL STRUCTURES FOR THE HYPERVARIABLE REGIONS OF IMMUNOGLOBULINS [J].

CHOTHIA, C ;

LESK, AM .

JOURNAL OF MOLECULAR BIOLOGY, 1987, 196 (04) :901-917

[4]

CONFORMATIONS OF IMMUNOGLOBULIN HYPERVARIABLE REGIONS [J].

CHOTHIA, C ;

LESK, AM ;

TRAMONTANO, A ;

LEVITT, M ;

SMITHGILL, SJ ;

AIR, G ;

SHERIFF, S ;

PADLAN, EA ;

DAVIES, D ;

TULIP, WR ;

COLMAN, PM ;

SPINELLI, S ;

ALZARI, PM ;

POLJAK, RJ .

NATURE, 1989, 342 (6252) :877-883

[5]

Dynamic transcriptome of Schizosaccharomyces pombe shown by RNA-DNA hybrid mapping [J].

Dutrow, Natalie ;

Nix, David A. ;

Holt, Derick ;

Milash, Brett ;

Dalley, Brian ;

Westbroek, Erick ;

Parnell, Timothy J. ;

Cairns, Bradley R. .

NATURE GENETICS, 2008, 40 (08) :977-986

[6]

Loss of Topoisomerase I leads to R-loop-mediated transcriptional blocks during ribosomal RNA synthesis [J].

El Hage, Aziz ;

French, Sarah L. ;

Beyer, Ann L. ;

Tollervey, David .

GENES & DEVELOPMENT, 2010, 24 (14) :1546-1558

[7]

R-loop-mediated genomic instability is caused by impairment of replication fork progression [J].

Gan, Wenjian ;

Guan, Zhishuang ;

Liu, Jie ;

Gui, Ting ;

Shen, Keng ;

Manley, James L. ;

Li, Xialu .

GENES & DEVELOPMENT, 2011, 25 (19) :2041-2056

[8]

R-Loop Formation Is a Distinctive Characteristic of Unmethylated Human CpG Island Promoters [J].

Ginno, Paul A. ;

Lott, Paul L. ;

Christensen, Holly C. ;

Korf, Ian ;

Chedin, Frederic .

MOLECULAR CELL, 2012, 45 (06) :814-825

[9]

Convergent transcription induces transcriptional gene silencing in fission yeast and mammalian cells [J].

Gullerova, Monika ;

Proudfoot, Nick J. .

NATURE STRUCTURAL & MOLECULAR BIOLOGY, 2012, 19 (11) :1193-+

[10]

An antibody-based microarray assay for small RNA detection [J].

Hu, Zonglin ;

Zhang, Aixia ;

Storz, Gisela ;

Gottesman, Susan ;

Leppla, Stephen H. .

NUCLEIC ACIDS RESEARCH, 2006, 34 (07)

← 1 2 3 →