Immunoaffinity techniques in analysis.
Immunoaffinity techniques in analysis.
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分析中的免疫亲和技术。
DOI：
10.1016/j.jchromb.2009.12.001
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Phillips,TerryM
中科院分区：
文献类型：
--
作者：
Phillips,TerryM
The use of antibodies as selection agents for isolating and measuring specific analytes has been reported since the 1960’s and led to the development of the classical immunoassay, wherein one antibody was used as a capture agent and a second antibody combined with a detectable label was used as the detection or reporter reagent. Following the development of the radioimmunoassay, the enzyme-linked immunoassay (ELISA) became one of the most popular immunological assays for measuring defined analytes in various fluids and tissues. However, neither of these assays allowed for the isolation of the analyte for further characterization or investigation, which led to a growing interest in applying the specificity of antibody-antigen reactions to the analytical and separation sciences. This interest focused on either developing immunoaffinity isolation procedures as an analytical technique in its own rights [1] or using antibody selectivity to isolate a specific analyte prior to further analysis. In the former case, the isolated analyte could be isolated and recovered, which is a distinct advantage over the ELISA technique in which the analyte is unrecoverable following the detection stage. In the latter case, the immunoaffinity step provided a specific cleanup prior to a more sophisticated analysis.A major advantage of antibody-based separation techniques is the specificity that antibodies bring to the separation, enabling a single analyte to be isolated from complex biological or chemical matrices. Additionally, antibodies can be assembled as a panel capable of isolating multiple analytes simultaneously, which can then be separated and analyzed by a second technique such as high-performance liquid chromatography (HPLC), capillary electrophoresis (CE), or capillary electrochromatography (CEC). The advent of engineered antibodies ie monoclonal, bifunctional, and single chain antibodies has made an impact on the popularity of immunoaffinity techniques in both chemistry and biology. These antibodies have lessened the necessity of extensive and often complicated purification of animal serum or plasma to produce class specific polyclonal antibodies, often containing multiple specificities. Further, the use of high affinity antibodies has greatly sped up the initial antibody capture phase, thus shortening the time of the analysis. The combination of immunoaffinity techniques with other analytical processes, especially mass spectrometry has made considerable impact on biomedical research especially proteomics and biomarker discovery [2, 3]. Immunoaffinity isolation has successfully been used in food analysis, toxicology [4] drug analysis and environmental monitoring [5].
