McInerney T L, Nice E, Jackson D C
Department of Microbiology, University of Melbourne, Parkville, Australia.
Biomed Pept Proteins Nucleic Acids. 1994;1(1):21-4.
The affinity of interaction between two monoclonal antibodies and a synthetic peptide representing the C-terminal 23 residues of the heavy chain (HA1) of influenza virus hemagglutinin were determined using an air-driven ultracentrifuge. The technique makes use of common laboratory equipment and is based on sound theoretical principles. Because the method does not rely on the solid-phase immobilisation of one of the interacting species, it circumvents problems associated with ELISA-like assays, which, in the case of peptides, may involve the immobilisation of ligand through association of amino acid residues necessary for recognition by antibody. The technique should be applicable to the study of a wide range of ligand-acceptor systems. Because only one of the reagents needs to be pure to allow labelling, prior purification of the biological receptor is not necessary. The method also lends itself to inhibition experiments in which the effects of various homologs on the binding event can be examined in a way which permits an evaluation of potential agonists and antagonists.
使用空气驱动超速离心机测定了两种单克隆抗体与代表流感病毒血凝素重链（HA1）C末端23个残基的合成肽之间的相互作用亲和力。该技术使用常见的实验室设备，且基于可靠的理论原理。由于该方法不依赖于相互作用物种之一的固相固定，它避免了与酶联免疫吸附测定（ELISA）类似的检测相关的问题，对于肽而言，ELISA可能涉及通过抗体识别所需的氨基酸残基缔合来固定配体。该技术应适用于广泛的配体-受体系统的研究。因为只需要一种试剂是纯的就能进行标记，所以无需事先纯化生物受体。该方法也适用于抑制实验，在抑制实验中，可以以允许评估潜在激动剂和拮抗剂的方式检查各种同源物对结合事件的影响。
