Sandwich Enzyme-Linked Immunosorbent Assays (SELISAs) Quantitate and Distinguish Two Forms of Transforming Growth Factor-Beta (TGF-beta 1 and TGF-beta 2) in Complex Biological Fluids

被引:127
作者
Danielpour, David [1 ]
Kim, Kyung-Young [1 ]
Dart, Linda L. [1 ]
Watanabe, Shinichi [1 ]
Roberts, Anita B. [1 ]
Sporn, Michael B. [1 ]
机构
[1] NCI, Lab Chemoprevent, Bldg 41,Rm C629, Bethesda, MD 20892 USA
关键词
transforming growth factor-beta; sandwich ELISA; turkey antibodies; rabbit antibodies; affinity purification; conditioned medium; trichloroacetic acid precipitation;
D O I
10.3109/08977198909069082
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
We have developed sandwich enzyme-linked immunosorbent assays (SELISAs) for TGF-beta 1 and TGF-beta 2 using both turkey and rabbit neutralizing polyclonal antibodies against native TGF-beta s. Each assay is based on the binding of two different antibodies to distinct epitopes of a single TGF-beta molecule. With these assays, TGF-beta types 1 and 2 can each be specifically quantitated in complex biological fluids, with detection limits of 2-5 pg. TGF-beta 3 and TGF-beta 5 either do not cross-react or cross-react very poorly in these assays. TGF-$1.2 heterodimer, although 50-80% neutralized by either TGF-beta 1 or TGF-beta 2 antibodies, shows only a 1.5 and 3.7% cross-reactivity in the TGF-beta 1 and TGF-beta 2 SELISAs, respectively. The SELISAs reported here represent the most specific, rapid, and precise assays for TGF-beta 1 and TGF-beta 2 reported thus far.
引用
收藏
页码:61 / 71
页数:11
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