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Deciphering Protein Corona by scFv-Based Affinity Chromatography.
Yuxiu Chu1, Wenjing Tang2, Zui Zhang1
1Department of Pharmacology, School of Basic Medical Sciences, and Center of Medical Research and Innovation, Shanghai Pudong Hospital, Fudan University, Shanghai, 200032, P.R. China.
Nano Letters
|February 22, 2021
Summary
Developing an anti-PEG single-chain variable fragment (PEG-scFv) affinity chromatography method precisely separates protein coronas on PEGylated liposomes. This technique enhances collection efficiency and preserves protein integrity for deeper analysis.
Area of Science:
- Biotechnology
- Nanomedicine
- Analytical Chemistry
Background:
- Precisely characterizing protein coronas on nanomedicines is challenging with traditional methods.
- Existing separation techniques like centrifugation suffer from low efficiency and contamination.
- Understanding protein corona composition is vital for nanomedicine efficacy and safety.
Purpose of the Study:
- To develop a novel affinity chromatography method for efficient and precise separation of protein coronas.
- To overcome limitations of traditional separation techniques for protein corona analysis.
- To enable deeper deciphering of protein corona structural and functional characteristics.
Main Methods:
- Development of an anti-PEG single-chain variable fragment (PEG-scFv) affinity chromatography (AfC) system.
- Utilizing His-tagged PEG-scFv to capture PEGylated liposomes (sLip) via a Ni-NTA column.
- Comparing AfC performance against traditional centrifugation for protein corona separation.
Main Results:
- AfC demonstrated a 43-fold higher protein corona collecting efficiency compared to centrifugation.
- AfC successfully separated sLip/protein complexes from plasma protein aggregates and endogenous vesicles.
- The AfC method preserved loosely bound proteins and evaded contamination, unlike centrifugation.
Conclusions:
- The developed scFv-based AfC provides an unprecedented approach for precise and efficient protein corona separation.
- This method is crucial for analyzing limited *in vivo* samples and offers improved resolution.
- scFv-based AfC opens new possibilities for studying protein coronas on diverse nanomedicines.
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