Greenfield Edward A
Cold Spring Harb Protoc. 2022 Jul 12;2022(7):Pdb.top100180. doi: 10.1101/pdb.top100180.
The traditional method for generating polyclonal and monoclonal antibodies requires the immunization of an animal. Selecting the best species of animal and getting that animal's immune system to respond to a target antigen with an antibody response are essential to obtaining good-quality antibodies and hybridomas. There are only a limited number of opportunities for a researcher to intervene to manipulate and tailor the response to a particular antigen. Here we present advice and methods for designing the way in which the antigen is presented to the immune system (i.e., the immunization protocol), including the choice of animal, the antigen dose, the use of adjuvants, the route and number of injections, and the period between injections.
传统的产生多克隆抗体和单克隆抗体的方法需要对动物进行免疫。选择最佳的动物物种并使该动物的免疫系统以抗体反应对靶抗原作出应答,对于获得高质量的抗体和杂交瘤至关重要。研究人员干预以操纵和调整对特定抗原的反应的机会有限。在此,我们提供有关设计将抗原呈递给免疫系统的方式(即免疫方案)的建议和方法,包括动物的选择、抗原剂量、佐剂的使用、注射途径和次数以及注射间隔时间。