Carlsson L, Mercado M, Baumann G, Stene M, Attie K, Reichert M, Albertsson-Wikland K, Dawson K, Wong W L
Research Centre for Endocrinology and Metabolism, University of Göteborg, Sweden.
Proc Soc Exp Biol Med. 1994 Jul;206(3):312-5. doi: 10.3181/00379727-206-43766.
The first method used for detection of growth hormone-binding protein (GHBP) in biological fluids was based on the incubation of the sample with radiolabeled GH followed by separation of bound and free GH by gel exclusion chromatography. Recently, other methods have been developed which are faster and easier to use. These methods include variants of the original binding/column assay (e.g., separation of bound and free GH is obtained by immunoprecipitation, charcoal adsorption, ion exchange chromatography, or HPLC), and a ligand-mediated immunofunctional assay (LIFA), in which a monoclonal antibody is used to capture the GHBP on a microtiter plate; all binding sites are saturated with GH and an anti-GH antibody is used to detect the amount of GH (endogenous and exogenous) bound to the GHBP. To permit comparison of results obtained by different methods we have cross-validated the LIFA with two different binding assays: (i) the original long column assay (column assay), and (ii) an assay based on immunoprecipitation (RIPA) of the GH/GHBP complex with an anti-GHBP antibody.
检测生物体液中生长激素结合蛋白（GHBP）的第一种方法是将样品与放射性标记的生长激素孵育，然后通过凝胶排阻色谱法分离结合型和游离型生长激素。最近，又开发出了其他更快且更易于使用的方法。这些方法包括原始结合/柱分析法的变体（例如，通过免疫沉淀、活性炭吸附、离子交换色谱法或高效液相色谱法分离结合型和游离型生长激素），以及一种配体介导的免疫功能分析法（LIFA），其中使用单克隆抗体在微量滴定板上捕获GHBP；所有结合位点都用生长激素饱和，并用抗生长激素抗体检测与GHBP结合的生长激素（内源性和外源性）的量。为了使不同方法得到的结果具有可比性，我们用两种不同的结合分析法对LIFA进行了交叉验证：（i）原始的长柱分析法（柱分析法），以及（ii）一种基于用抗GHBP抗体对GH/GHBP复合物进行免疫沉淀（RIPA）的分析法。
