Short peptide biomimetic chromatography technology as a developing protein separation technology has huge potential for antibody purification. In this study, four tetrapeptide ligands (Ac-FYKH, Ac-YEHF, Ac-YFLH and Ac-FYHI) with high potential binding ability to antibody were selected for the optimal ligand to antibody purification. The results showed that Ac-YEHF-4FF resin had higher binding capacity and selectivity for hIgG among the four resins. And at pH 7.0 and 0.3 ml/min, the highest Q_10%-hIgG of Ac-YEHF-4FF resin was 26.2 mg/ml resin while its Q_10%-BSA was just 2.2 mg/ml resin. Further, Ac-YEHF-4FF resin was used to purify protein mixtures. By binding at pH 7.0 and being eluted at pH 5.0 and pH 4.0, Ac-YEHF-4FF resin was well used to separate hIgG from BSA containing feedstock, HSA containing feedstock and human serum with the purity and yield both more than 95%. And the screened resin could also separate mAb from CHO cell culture supernatant with purity 94.3% and yield 97.5%. The adsorption and separation results of Ac-YEHF-4FF resin indicated that the goal of getting the efficacy of critical residues from protein A to biomimetic its structure and function could be achieved, which had great significance to the establishment and improvement of tetrapeptide biomimetic chromatography, and also provided a new method for the field of antibody separation and purification.

短肽仿生层析技术作为一种发展中的蛋白质分离技术，在抗体纯化方面具有巨大的潜力。在本研究中，选择了与抗体具有高潜在结合能力的四种四肽配体（Ac-FYKH、Ac-YEHF、Ac-YFLH 和 Ac-FYHI）作为抗体纯化的最佳配体。结果表明Ac-YEHF-4FF树脂在四种树脂中对hIgG的结合能力和选择性均较高。在 pH 7.0 和 0.3 ml/min 时，Ac-YEHF-4FF 树脂的最高Q _10%-hIgG为 26.2 mg/ml 树脂，而其Q _10%-BSA仅为 2.2 mg/ml 树脂。此外，Ac-YEHF-4FF 树脂用于纯化蛋白质混合物。通过在 pH 7.0 下结合并在 pH 5.0 和 pH 4.0 下洗脱，Ac-YEHF-4FF 树脂很好地用于将 hIgG 从含 BSA 的原料、含 HSA 的原料和人血清中分离出来，纯度和收率均超过 95%。并且筛选出的树脂还可以从CHO细胞培养上清液中分离出mAb，纯度为94.3%，收率97.5%。Ac-YEHF-4FF树脂的吸附分离结果表明，获得蛋白质A关键残基的功效以仿生其结构和功能的目标可以实现，这对四肽仿生层析的建立和改进具有重要意义。也为抗体分离纯化领域提供了一种新的方法。