How eluents define proteomic fingerprinting of protein corona on nanoparticles

Qiu, Liangjia1; Zhang, Ying2,3; Wei, Genxia4; Wang, Chen2,3; Zhu, Yinhua2,3; Yang, Tong2,3; Chu, Zheng2,3; Gao, Peng1; Cheng, Guangqing2,3; Ma, Ang2,3; Wong, Yin Kwan5; Zhang, Junzhe2,3; Xu, Chengchao2,3; Wang, Jigang1,2,3,6,7; Tang, Huan2,3

2023-10-15

10.1016/j.jcis.2023.05.045

JOURNAL OF COLLOID AND INTERFACE SCIENCE   影响因子和分区

0021-9797

1095-7103

Nanoparticles (NPs) have broad application prospects in the field of biomedicine due to their excellent physi-cochemical properties. When entering biological fluids, NPs inevitably encountered proteins and were subse-quently surrounded by them, forming the termed protein corona (PC). As PC has been evidenced to have critical roles in deciding the biological fates of NPs, how to precisely characterize PC is vital to promote the clinical translation of nanomedicine by understanding and harnessing NPs' behaviors. During the centrifugation-based separation techniques for the PC preparation, direct elution has been most widely used to strip proteins from NPs due to its simpleness and robustness, but the roles of multifarious eluents have never been systematically declared. Herein, seven eluents composed of three denaturants, sodium dodecyl sulfate (SDS), dithiothreitol (DTT), and urea (Urea), were applied to detach PC from gold nanoparticles (AuNPs) and silica nanoparticles (SiNPs), and eluted proteins in PC have been carefully characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and chromatography coupled tandem mass spectrometry (LC-MS/MS). Our re-sults showed that SDS and DTT were the main contributors to the efficient desorption of PC on SiNPs and AuNPs, respectively. The molecular reactions between NPs and proteins were explored and verified by SDS-PAGE analysis of PC formed in the serums pretreated with protein denaturing or alkylating agents. The proteomic fingerprinting analysis indicated the difference of the eluted proteins brought by the seven eluents was the abundance rather than the species. The enrichment of some opsonins and dysopsonins in a special elution re-minds us that the possibility of biased judgments on predicting NPs' biological behaviors under different elution conditions. The synergistic effects or antagonistic effects among denaturants for eluting PC were manifested in a nanoparticle-type dependent way by integrating the properties of the eluted proteins. Collectively, this study not only underlines the urgent need of choosing the appropriate eluents for identifying PC robustly and unbiasedly, but also provides an insight into the understanding of molecular interactions during PC formation.

Protein corona Proteomics Nanoparticles Elution Nano-bio interface

SCI

英语

通讯

National Key Research and Development Program of China[2020YFA0908000] ; Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine[ZYYCXTD-C-202002] ; National Natural Science Foundation of China["32201177","82141001","82074098"] ; CACMS Innovation Fund["CI2021A05101","CI2021A05104"] ; Fundamental Research Funds for the Central public welfare research institutes["ZZ14-YQ-061","ZZ16-ND-10-24","ZZ14-ND-010","ZZ15-ND-10","ZZ14-FL-002","ZZ16-ND-10-27"] ; null[ZZ14-YQ-057]

Chemistry

Chemistry, Physical

WOS:001023836000001

ACADEMIC PRESS INC ELSEVIER SCIENCE

CHEMISTRY

Web of Science

1.Southern Med Univ, Sch Pharmaceut Sci, Guangdong Key Lab New Drug Screening, Guangzhou 510515, Guangdong, Peoples R China
2.China Acad Chinese Med Sci, Artemisinin Res Ctr, Beijing 100700, Peoples R China
3.China Acad Chinese Med Sci, Inst Chinese Mat Med, Beijing 100700, Peoples R China
4.Southern Med Univ, Nanfang Hosp, Huiqiao Med Ctr, Guangzhou 510515, Peoples R China
5.Natl Univ Singapore, Dept Physiol Sci, Singapore 117543, Singapore
6.Southern Univ Sci & Technol, Shenzhen Key Lab Kidney Dis, Dept Nephrol, Shenzhen 518020, Peoples R China
7.Southern Univ Sci & Technol, Jinan Univ, Shenzhen Peoples Hosp, Affiliated Hosp 1,Shenzhen Clin Res Ctr Geriatr,Cl, Shenzhen 518020, Peoples R China

Qiu, Liangjia,Zhang, Ying,Wei, Genxia,et al. How eluents define proteomic fingerprinting of protein corona on nanoparticles[J]. JOURNAL OF COLLOID AND INTERFACE SCIENCE,2023,648.

Qiu, Liangjia.,Zhang, Ying.,Wei, Genxia.,Wang, Chen.,Zhu, Yinhua.,...&Tang, Huan.(2023).How eluents define proteomic fingerprinting of protein corona on nanoparticles.JOURNAL OF COLLOID AND INTERFACE SCIENCE,648.

Qiu, Liangjia,et al."How eluents define proteomic fingerprinting of protein corona on nanoparticles".JOURNAL OF COLLOID AND INTERFACE SCIENCE 648(2023).