Antibody isoelectric spectra. Analysis of the heterogeneity of antibody molecules in serum by isoelectric focusing in gel and specific detection with hapten
Antibody isoelectric spectra. Analysis of the heterogeneity of antibody molecules in serum by isoelectric focusing in gel and specific detection with hapten
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抗体等电谱。
DOI：
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发表时间：
1971
影响因子：
5.4
通讯作者：
A. Williamson
中科院分区：
文献类型：
--
作者：
A. Williamson
Analytical isoelectric focusing in thin layers of polyacrylamide gel was shown by Awdeh, Williamson & Askonas (Nature 1968. 219: 66) to be capable of resolving a specifically purified antibody into a pattern of discrete bands; this has been called the antibody isoelectric spectrum. The usefulness of this method has been extended by employing radioactive hapten to visualize the isoelectric spectrum of anti‐hapten antibodies present in sera. Samples of immune serum are focused, usually using pH 5–8 range Ampholine. Labeled hapten is applied to the surface of the thin layer gel and allowed to combine with focused antibodies; after fixation of the antibody‐hapten complexes, and washing to remove excess hapten, the gel is air dried and a radioautograph taken on X‐ray film. This visualization technique has been shown to be highly specific for antibody against the developing hapten. The sensitivity of detection of antibody is primarily limited only by the specific activity of the radioactive hapten. The visualization of anti‐DNP spectra is described and the extension of the method to other antibodies is discussed. An interpretation of the isoelectric spectrum of antibody present in serum in terms of the number of different antibodies originally synthesized by the animal is presented.
