abstract
Although pentraxin-3 holds promise as a diagnosis/prognosis biomarker of microbial infections and lung cancer, its analysis in human serum can be constrained by matrix effects caused by high abundance proteins - human serum albumin and immunoglobulin G. Aqueous biphasic systems composed of polymers and citrate buffer are here proposed as a serum pretreatment step to improve the accuracy of pentraxin-3 analysis. Binodal curves were determined to identify the compositions required to form two phases and to correlate the polymers' properties and performance in serum pretreatment and biomarker extraction. Aqueous biphasic systems were evaluated regarding their ability to deplete human serum albumin and immunoglobulin G at the interphase. Polymers of relatively high to intermediate hydrophobicity were unveiled as efficient components to deplete high abundance serum proteins. Considering the possibility to extract pentraxin-3 from human serum into the polymer-rich phase, the system composed of polyethylene glycol with a molecular weight of 1000 g & sdot;mol(- 1) simultaneously achieved >93 % of human serum albumin and immunoglobulin G depletion and complete biomarker extraction. The accuracy of analysis of pretreated human serum by enzyme-linked immunosorbent assays outperformed that of a non-pretreated sample, with a relative error of 0.8 % compared to 14.6 %, contributing to boost pentraxin-3 usefulness as a biomarker.
keywords
LIQUID-LIQUID EQUILIBRIA; 2-PHASE SYSTEMS; POLYETHYLENE-GLYCOL; POLYPROPYLENE GLYCOL; IMMUNOGLOBULIN-G; INNATE IMMUNITY; PROTEIN PRECIPITATION; PURIFICATION; EXTRACTION; WATER
subject category
Biochemistry & Molecular Biology; Chemistry; Polymer Science
authors
Mendes, MSM; Rosa, ME; Coutinho, JAP; Freire, MG; Silva, FAE
our authors
Ana Francisca Osório de Almeida Coelho e Silva
Junior Researcher
Mara Guadalupe Freire Martins
Coordinating ResearcherGroups
G4 - Renewable Materials and Circular Economy
G5 - Biomimetic, Biological and Living Materials
Projects
CICECO - Aveiro Institute of Materials (UIDB/50011/2020)
CICECO - Aveiro Institute of Materials (UIDP/50011/2020)
Associated Laboratory CICECO-Aveiro Institute of Materials (LA/P/0006/2020)
Collaboratory for Emerging Technologies, CoLab (EMERGING TECHNOLOGIES)
acknowledgements
This work was developed within the scope of the project CICECO-Aveiro Institute of Materials, UIDB/50011/2020, UIDP/50011/2020 & LA/P/0006/2020, financed by national funds through the FCT/MEC (PIDDAC). This work was developed within the project PTDC/EMD-TLM/3253/2020 (ILSurvive), funded by national funds (OE), through FCT/MCTES. M.S.M.M. and M.E.R. acknowledge FCT for the doctoral grants 2022.11229.BD and SFRH/BD/136995/2018, respectively. F.A. eS. acknowledges FCT for the researcher contract CEECIND/03076/2018 under the Scientific Employment Stimulus - Individual Call 2018.