abstract
Analysing biomarkers in human serum could be used as an effective and less invasive approach for the diagnosis of lung cancer; however, biomarker detection reliability is highly limited due to matrix effects. Herein, aqueous biphasic systems (ABS) are studied as platforms for human serum pretreatment, allowing the simultaneous depletion of high abundant proteins and biomarker extraction. By using ABS varying the polyethylene glycol (PEG) molecular weight between 400 and 6000 g center dot mol- 1 and adopting phosphate buffer as the other phaseforming component, the depletion of the high abundance serum proteins immunoglobulin G (IgG) and human serum albumin (HSA) is induced at the ABS interphase, through precipitation, forming a three-phase partitioning system (ABS-TPP). Maximum depletion efficiencies of 99 % for IgG and 70 % for HSA were achieved in one step using PEG 1500-based ABS-TPP. On the other hand, lung cancer biomarkers, such as CYFRA 21-1, are extracted to the PEG-rich phase of the same ABS-TPP with recovery yields of 91 %. This work shows that a proper selection of the PEG molecular weight in the ABS composition leads to the efficient depletion of high-abundance proteins and extraction of cancer biomarkers from human serum, in a single step, confirming the potential of ABS for sample pretreatment to improve biomarker analysis.
keywords
2-PHASE SYSTEMS; POLYETHYLENE-GLYCOL; PURIFICATION; PRECIPITATION; ALBUMIN; PEG; INTERFERENCE; RARE; IGG; DNA
subject category
Chemistry
authors
Rosa, ME; Mendes, MSM; Belchior, DCV; Coutinho, JAP; Silva, FAE; Freire, MG
our authors
Ana Francisca Osório de Almeida Coelho e Silva
Junior Researcher
Mara Guadalupe Freire Martins
Coordinating Researcher
Marguerita Evangelho da Rosa
PhD StudentGroups
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 (DOI 10.54499/UIDB/50011/2020) , UIDP/50011/2020 (DOI 10.54499/UIDP/50011/2020) & LA/P/0006/2020 (DOI 10.54499/LA/P/0006/2020) , financed by national funds through the FCT/MCTES (PIDDAC) . This work was developed within the project ILSurvive, PTDC/EMD-TLM/3253/2020 (DOI 10.54499/PTDC/EMD-TLM/3253/2020) , 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/CP1559/CT0024 (DOI 10.54499/CEE-CIND/03076/2018/CP1559/CT0024) under the Scientific Employment Stimulus - Individual Call 2018.