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

被引：333

作者：

BOGUSLAWSKI, SJ ^{[1
]}

SMITH, DE ^{[1
]}

MICHALAK, MA ^{[1
]}

MICKELSON, KE ^{[1
]}

YEHLE, CO ^{[1
]}

PATTERSON, WL ^{[1
]}

CARRICO, RJ ^{[1
]}

机构：

[1] MILES LABS INC, AMES DIV, POB 70, ELKHART, IN 46515 USA

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 1986年 / 89卷 / 01期

关键词：

D O I：

10.1016/0022-1759(86)90040-2

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Monoclonal antibodies were raised to a DNA .cntdot. RNA heteropolymer duplex prepared by transcription of .PHI.X174 single-stranded DNA with DNA-dependent RNA polymerase. A monoclonal antibody with the highest affinity and specificity was selected. This antibody bound the DNA .cntdot. RNA heteropolymer and poly(I) .cntdot. poly(dC) equally but 100-fold higher levels of poly(A) .cntdot. poly(dT) were required to achieve a similar degree of binding in competitive binding assays using DNA .cntdot. [3H]RNA. Single-stranded DNA, double-stranded DNA and RNA, and ribosomal RNA were not bound by the antibody. The observed association constant for the antibody and DNA .cntdot. [3H]RNA, determined by Scatchard analysis, was 8.5 .times. 1010 l/mol assuming independent antibody binding sites. The antibody and an alkaline phosphatase-labeled second antibody were used in an immunodetection method for measurement of hybrids formed between immobilized DNA probes of various lengths and 23 S ribosomal RNA. The colorimetric response of this assay increased linearly with the amount of hybrid formed.

引用

页码：123 / 130

页数：8

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