Delahaut Philippe
CER Groupe, rue du Point du Jour 8, 6900 Marloie, Belgium.
Methods. 2017 Mar 1;116:4-11. doi: 10.1016/j.ymeth.2017.01.002. Epub 2017 Jan 10.
Polyclonal antibodies are frequently used as immunodiagnostic tools in fundamental research. They are also used for routine diagnostic purposes in human and veterinary medicine and for quality control procedures in the food-processing industry. The antibody is a major component of the detection system. It binds with the molecule to be identified. This conjugate is subsequently revealed by means of binding the antibody with a radio-isotope, a fluorescent substance, an enzyme inducing a color change, or a biosensor based analytical system. Polyclonal antibodies are also used for treatment purposes in various pathologies. They might have immunomodulating or anti-inflammatory properties. Snake venom, rabies and tetanus antisera are examples of a therapeutic application; immunosuppressive antithymocyte serum used in order to avoid rejection in organ transplantation is another example from human medicine. These therapeutic aids need hyperimmunisation of animals. Since these are subject to a certain number of interventions such as injections and blood samplings, animal welfare prescriptions have to be taken into account. The optimisation of the immunisation protocol allows for reducing the numbers of animals used as well as reducing stress and pain while obtaining high quality antibodies. This article describes the critical steps in polyclonal antibody production with a particular focus on the choice of the animal species, the age of the subjects, the injection protocol and the sampling times.
多克隆抗体在基础研究中经常被用作免疫诊断工具。它们也用于人类和兽医学的常规诊断目的以及食品加工业的质量控制程序。抗体是检测系统的主要组成部分。它与待识别的分子结合。随后,通过将抗体与放射性同位素、荧光物质、能引起颜色变化的酶或基于生物传感器的分析系统结合来揭示这种结合物。多克隆抗体也用于各种病症的治疗目的。它们可能具有免疫调节或抗炎特性。蛇毒、狂犬病和破伤风抗血清是治疗应用的例子;为避免器官移植排斥而使用的免疫抑制抗胸腺细胞血清是人类医学中的另一个例子。这些治疗辅助手段需要对动物进行超免疫。由于这些动物要接受一定数量的干预,如注射和采血,所以必须考虑动物福利规定。优化免疫方案可以减少所用动物的数量,同时在获得高质量抗体的同时减轻压力和疼痛。本文描述了多克隆抗体制备中的关键步骤,特别关注动物种类的选择、实验对象的年龄、注射方案和采样时间。