Effective Bioactivity Retention of Low-Concentration Antibodies on HFBI-Modified Fluorescence ICTS for Sensitive and Rapid Detection of PSA.
Effective Bioactivity Retention of Low-Concentration Antibodies on HFBI-Modified Fluorescence ICTS for Sensitive and Rapid Detection of PSA.
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DOI：
10.1021/acsami.8b02945
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发表时间：
2018-04
影响因子：
9.5
通讯作者：
Bo Zhang;Weichen Gao;Jiafang Piao;Yunjie Xiao;Bin Wang;Weipan Peng;Xiaoqun Gong;Zefang Wang;Haitao Yang;Jin Chang
中科院分区：
文献类型：
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作者：
Bo Zhang;Weichen Gao;Jiafang Piao;Yunjie Xiao;Bin Wang;Weipan Peng;Xiaoqun Gong;Zefang Wang;Haitao Yang;Jin Chang
如今，提高分析灵敏度仍然是构建膜基荧光免疫层析测试条（FICTS）的一大挑战。然而，对于直接影响分析灵敏度的多孔硝酸纤维素膜（PNM）测试线（T线）上固定的抗体（Ab）的生物活性研究较少。在这项工作中，引入了一种新型两亲疏水蛋白（HFBI）蛋白来修饰 T 系，以有效保留 Abs 的生物活性。结果表明，HFBI 可以在 PNM 上自组装，并将 Abs 固定在“直立”方向。与传统FICTS相比，T线上仅含有0.2 mg/mL单克隆抗体的HFBI改良FICTS能够实现更准确的定量检测和更好的灵敏度（前列腺特异性抗原为0.06 ng/mL），比T线上相同浓度单克隆抗体的传统FICTS低2个数量级以上。此外，通过检测150份临床血清样本，考察了HFBI改进的FICTS的准确性，检测结果与电化学发光免疫分析结果一致。我们的结果为 FICTS 上的抗体固定化提供了一种新颖且有前景的策略，适用于患者附近和护理点应用。
Nowadays, increasing analytical sensitivity is still a big challenge in constructing membrane-based fluorescence immunochromatography test strips (FICTS). However, the bioactivity of antibody (Ab) immobilized on the test line (T line) of porous nitrocellulose membrane (PNM), which directly influences the analytical sensitivity, is less studied. In this work, a novel amphiphilic hydrophobin (HFBI) protein was introduced to modify the T line to effectively retain the Abs' bioactivity. The results indicated that HFBI could self-assemble on the PNM and immobilize the Abs in the "stand-up" orientation. Compared with the conventional FICTS, the HFBI-modified FICTS with only 0.2 mg/mL of monoclonal Abs on T line enable more accurate quantitative detection and better sensitivity (0.06 ng/mL for prostate specific antigen), which is more than 2 orders of magnitude lower than that of the conventional FICTS with the same concentration of monoclonal Abs on T line. Furthermore, the accuracy of this HFBI-modified FICTS was investigated by testing 150 clinical serum samples and the detection results were coincident with those by electrochemiluminescence immunoassay. Our results provide a novel and promising strategy of Ab immobilization on FICTS for near-patient and point-of-care application.