Luna Omar F, Perez Yomkippur V, Ferrari Daniele P, Sayedipour Sana S, Royo Miriam, Acosta Gerardo A, Cruz Luis J, Alves Frauke, Agner Erik, Sydnes Magne O, Albericio Fernando
Department of Organic Chemistry, University of Barcelona, and CIBER-BBN, Networking Centre on Bioengineering, Biomaterials, and Nanomedicine, Barcelona 08028, Spain.
Polypure AS, Martin Linges vei 25, Fornebu 1364, Norway.
ACS Omega. 2024 Aug 1;9(32):34544-34554. doi: 10.1021/acsomega.4c02604. eCollection 2024 Aug 13.
Peptide-based cancer vaccines have shown promising results in preclinical trials focusing on tumor immunotherapy. However, the presence of hydrophobic amino acid segments within these peptide sequences poses challenges in their synthesis, purification, and solubility, thereby hindering their potential use as cancer vaccines. In this study, we successfully synthesized peptide sequences derived from mesothelin (MSLN), a tumor-associated antigen overexpressed in pancreatic ductal adenocarcinoma (PDAC) by conjugating them with monodisperse polyethylene glycol (PEG). By PEGylating mesothelin epitopes of varying lengths (ranging from 9 to 38 amino acids) and hydrophobicity (60-90%), we achieved an effective method to improve the peptide yield and facilitate the processes of synthesis and purification. PEGylation significantly enhanced the solubility, facilitating the single-step purification of lengthy hydrophobic peptides. Most importantly, PEGylation did not compromise cell viability and had little to no effect on the immunogenicity of the peptides. In contrast, the addition of a palmitoyl group to increase immunogenicity led to reduced yield and solubility. Overall, PEGylation proves to be an effective technique for enhancing the solubility and broadening the range of utility of diverse long hydrophobic peptides.
基于肽的癌症疫苗在专注于肿瘤免疫疗法的临床前试验中已显示出有前景的结果。然而，这些肽序列中存在疏水性氨基酸片段，在其合成、纯化和溶解性方面带来了挑战，从而阻碍了它们作为癌症疫苗的潜在应用。在本研究中，我们通过将源自间皮素（MSLN）的肽序列与单分散聚乙二醇（PEG）偶联，成功合成了该肽序列，间皮素是一种在胰腺导管腺癌（PDAC）中过表达的肿瘤相关抗原。通过对不同长度（9至38个氨基酸）和疏水性（60 - 90%）的间皮素表位进行聚乙二醇化，我们实现了一种有效方法来提高肽产量并促进合成和纯化过程。聚乙二醇化显著增强了溶解性，便于对长链疏水肽进行单步纯化。最重要的是，聚乙二醇化不会损害细胞活力，并且对肽的免疫原性几乎没有影响。相比之下，添加棕榈酰基以增加免疫原性会导致产量和溶解性降低。总体而言，聚乙二醇化被证明是一种有效技术，可增强溶解性并拓宽各种长链疏水肽的应用范围。
