Effect of the Protein Corona on Antibody–Antigen Binding in Nanoparticle Sandwich Immunoassays
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- 作者：Helena de Puig ; Irene Bosch ; Marc Carré-Camps ; Kimberly Hamad-Schifferli
- 刊名：Bioconjugate Chemistry
- 出版年：2017
- 出版时间：January 18, 2017
- 年：2017
- 卷：28
- 期：1
- 页码：230-238
- 全文大小：474K
- ISSN：1520-4812
文摘
We investigated the effect of the protein corona on the function of nanoparticle (NP) antibody (Ab) conjugates in dipstick sandwich immunoassays. Ab specific for Zika virus nonstructural protein 1 (NS1) were conjugated to gold NPs, and another anti-NS1 Ab was immobilized onto the nitrocellulose membrane. Sandwich immunoassay formation was influenced by whether the strip was run in corona forming conditions, i.e., in human serum. Strips run in buffer or pure solutions of bovine serum albumin exhibited false positives, but those run in human serum did not. Serum pretreatment of the nitrocellulose also eliminated false positives. Corona formation around the NP-Ab in serum was faster than the immunoassay time scale. Langmuir binding analysis determined how the immobilized Ab affinity for the NP-Ab/NS1 was impacted by corona formation conditions, quantified as an effective dissociation constant, KDeff. Results show that corona formation mediates the specificity and sensitivity of the antibody–antigen interaction of Zika biomarkers in immunoassays, and plays a critical but beneficial role.