The utilization of immuno-magnetic nanoparticles (MNPs) for the selective capture, enrichment, and separation of specific glycoproteins from complicated biological samples is appealing for the discovery of disease biomarkers. Herein, MNPs were designed and anchored with abundant boronic acid (BA) and photoreactive alkyl diazirine (Diaz) functional groups to obtain permanently tethered Fc-fused Siglec-2 and antiserum amyloid A (SAA) mAb with the assistance of reversible boronate affinity and UV light activation in an orientation-controlled manner. The Siglec-2–Fc-functionalized MNPs showed excellent stability in fetal bovine serum (FBS) and excellent efficiency in the extraction of cell membrane glycoproteins. The anti-SAA mAb-functionalized MNPs maintained active Ab orientation and preserved antigen recognition capability in biological samples. Thus, the BA–Diaz-based strategy holds promise for the immobilization of glycoproteins, such as antibodies, with the original protein binding activity maintained, which can provide better enrichment for the sensitive detection of target proteins.

中文翻译：

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基于硼酸亲和力的光活化磁性纳米粒子，用于 Fc 融合凝集素和抗体的定向和不可逆缀合


利用免疫磁性纳米粒子（MNP）从复杂的生物样品中选择性捕获、富集和分离特定糖蛋白对于疾病生物标志物的发现很有吸引力。在此，设计了 MNP，并用丰富的硼酸 (BA) 和光反应性烷基二氮丙啶 (Diaz) 官能团进行锚定，以在可逆硼酸亲和力和 UV 的帮助下获得永久束缚的 Fc 融合的 Siglec-2 和抗血清淀粉样蛋白 A (SAA) mAb以方向控制的方式激活光。 Siglec-2–Fc 功能化的 MNP 在胎牛血清 (FBS) 中表现出优异的稳定性，并且在细胞膜糖蛋白的提取中表现出优异的效率。抗SAA mAb功能化的MNP在生物样品中保持了活性Ab方向并保留了抗原识别能力。因此，基于BA-Diaz的策略有望固定糖蛋白（例如抗体），同时保持原始的蛋白质结合活性，从而可以为目标蛋白的灵敏检测提供更好的富集。