Department of Pharmaceutical Chemistry, University of California, San Francisco, United States.
Department of Biochemistry and Biophysics, University of California, San Francisco, United States.
Curr Opin Biotechnol. 2019 Dec;60:153-158. doi: 10.1016/j.copbio.2019.01.012. Epub 2019 Mar 5.
Antibodies (Abs) are ubiquitous reagents for biological and biochemical research and are rapidly expanding into new therapeutic areas. They are one of the most important probes for determining how proteins function under normal and pathophysiological conditions. Abs are required for the quantification of targets, detection of temporal and spatial patterns of protein expression in cells and tissues, and identification of interacting partners and their biological activities. Their remarkable specificity and unique binding properties can facilitate three-dimensional structure determination using X-ray crystallography and electron cryomicroscopy. While hybridoma technology that involves animal immunization is often productive, many antigen targets do not generate useful Abs. This is particularly true if unique states of the target or critical non-immunogenic target sequences need to be recognized by the Abs. By using the methods of recombinant antibody generation, identification, and engineering, these 'hybridoma-refractory' antigens can be readily targeted. Specific, reproducible, and renewable recombinant Abs are proving to be invaluable reagents in applications ranging from biological discovery to structure determination of challenging macromolecules.
抗体(Abs)是生物和生化研究中无处不在的试剂,并且正在迅速扩展到新的治疗领域。它们是确定蛋白质在正常和病理生理条件下如何发挥作用的最重要探针之一。Abs 用于定量靶标,检测细胞和组织中蛋白质表达的时空模式,并鉴定相互作用的伙伴及其生物学活性。它们出色的特异性和独特的结合特性可以使用 X 射线晶体学和电子 cryomicroscopy 促进三维结构的确定。尽管涉及动物免疫的杂交瘤技术通常具有生产力,但许多抗原靶标不会产生有用的 Abs。如果需要 Abs 识别靶标的独特状态或关键非免疫原性靶序列,则尤其如此。通过使用重组抗体生成、鉴定和工程的方法,可以轻松地针对这些“杂交瘤难治性”抗原。特异性、可重现性和可再生的重组 Abs 正在证明在从生物发现到具有挑战性的大分子结构确定的各种应用中是非常有价值的试剂。