Eureka Therapeutics, Emeryville, CA, USA.
Department of Pediatrics, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
Methods Mol Biol. 2023;2702:327-345. doi: 10.1007/978-1-0716-3381-6_17.
Antibodies that bind peptide-MHC (pMHC) complex in a manner akin to T cell receptor (TCR) have not only helped in understanding the mechanism of TCR-pMHC interactions in the context of T cell biology but also spurred considerable interest in recent years as potential cancer therapeutics. Traditional methods to generate such antibodies using hybridoma and B cell sorting technologies are sometimes inadequate, possibly due to the small contribution of peptide to the overall B cell epitope space on the surface of the pMHC complex (typical peptide MW = 1 kDa versus MHC MW = 45 kDa) and to the multiple efficiency limiting steps inherent in these methods. In this chapter we describe phage display approaches, including a cell panning strategy, for the rapid generation of such antibodies with high specificity and affinity.
与 T 细胞受体 (TCR) 类似的方式结合肽-MHC (pMHC) 复合物的抗体不仅有助于理解 TCR-pMHC 相互作用在 T 细胞生物学背景下的机制，而且近年来作为潜在的癌症治疗方法引起了相当大的兴趣。使用杂交瘤和 B 细胞分选技术生成此类抗体的传统方法有时不够充分，这可能是由于肽对 pMHC 复合物表面总体 B 细胞表位空间的贡献较小（典型的肽 MW=1kDa 与 MHC MW=45kDa），以及这些方法固有的多个效率限制步骤。在本章中，我们描述了噬菌体展示方法，包括一种细胞淘选策略，用于快速生成具有高特异性和亲和力的此类抗体。
