PMID- 35033252
OWN - NLM
STAT- MEDLINE
DCOM- 20220118
LR  - 20220118
IS  - 1873-4324 (Electronic)
IS  - 0003-2670 (Linking)
VI  - 1191
DP  - 2022 Jan 25
TI  - A linear-polymer-based lactoferrin-selective recognition element for an ELISA 
      mimic: A proof of concept.
PG  - 339309
LID - S0003-2670(21)01135-1 [pii]
LID - 10.1016/j.aca.2021.339309 [doi]
AB  - The synthesis of polymers with tailored properties for the recognition of 
      macromolecules such as proteins is challenging. In this work, the synthesis of a 
      new polymer format, a linear polymer (LP), as the selective recognition element 
      for the globular protein lactoferrin (LF) is proposed as a proof-of-concept 
      study. For the synthesis, a solid-phase strategy using the reversible 
      deactivation radical polymerisation (RDRP) mechanism is proposed. This approach, 
      which is usually used in molecular imprinting, involves the immobilisation of LF 
      on the surface of a solid support, but, unlike classical imprinting, a 
      cross-linker in the polymerisation mixture is not required. Consequently, the 
      copolymer is soluble and flexible, thus overcoming the drawbacks associated with 
      traditional synthetic polymers for macromolecule imprinting. This new polymer 
      format has great potential for replacing natural antibodies in bioassays such as 
      enzyme-linked immunosorbent assays (ELISA), dot blot, western blot, or pull-down. 
      In our case, the linear polymer was used as a recognition element to replace 
      natural antibodies in a LF-selective ELISA. The responses of the linear polymer 
      between LF concentrations of 0.1 nM and 0.25 muM were studied, and a significant 
      difference was observed between the non-specific signals and the signals measured 
      in the presence of the polymeric material. Further, the response versus log 
      concentration curves were fitted to a logistic equation, allowing estimation of 
      the EC(50) value: 11.8 +/- 1.4 nM. We also confirmed the selective detection of LF 
      using the competitive inhibition of the selective LF-biotin conjugate (LF-Bi) 
      binding to the plastic receptor (LP) for closely related proteins (e.g. those 
      having similar molecular weights or isoelectric points) such as human lysozyme, 
      trypsin, and albumin, which are present in human body fluids. The system presents 
      a cross-reactivity value or selectivity of 1.95% for lysozyme, 0.028% for 
      trypsin, and 0.016% for albumin. The applicability of this method for the 
      determination of urine LF levels in inflammatory and infectious diseases of the 
      human urinary tract is also demonstrated.
CI  - Copyright (c) 2021. Published by Elsevier B.V.
FAU - Goicolea, M A
AU  - Goicolea MA
AD  - Department of Analytical Chemistry, Faculty of Pharmacy, University of the Basque 
      Country (UPV/EHU), 01006, Vitoria-Gasteiz, Spain. Electronic address: 
      mariaaranzazu.goicolea@ehu.eus.
FAU - Gomez-Caballero, A
AU  - Gomez-Caballero A
AD  - Department of Analytical Chemistry, Faculty of Pharmacy, University of the Basque 
      Country (UPV/EHU), 01006, Vitoria-Gasteiz, Spain.
FAU - Saumell-Esnaola, M
AU  - Saumell-Esnaola M
AD  - Department of Pharmacology, Faculty of Pharmacy, University of the Basque Country 
      (UPV/EHU), 01006, Vitoria-Gasteiz, Spain; Instituto de Investigacion Sanitaria 
      Bioaraba, Neurofarmacologia Celular y Molecular, 01008, Vitoria-Gasteiz, Spain.
FAU - Garcia Del Cano, G
AU  - Garcia Del Cano G
AD  - Department of Neurosciences, Faculty of Pharmacy, University of the Basque 
      Country (UPV/EHU), 01006, Vitoria-Gasteiz, Spain; Instituto de Investigacion 
      Sanitaria Bioaraba, Neurofarmacologia Celular y Molecular, 01008, 
      Vitoria-Gasteiz, Spain.
FAU - Unceta, N
AU  - Unceta N
AD  - Department of Analytical Chemistry, Faculty of Pharmacy, University of the Basque 
      Country (UPV/EHU), 01006, Vitoria-Gasteiz, Spain.
FAU - Salles, J
AU  - Salles J
AD  - Department of Pharmacology, Faculty of Pharmacy, University of the Basque Country 
      (UPV/EHU), 01006, Vitoria-Gasteiz, Spain; Instituto de Investigacion Sanitaria 
      Bioaraba, Neurofarmacologia Celular y Molecular, 01008, Vitoria-Gasteiz, Spain; 
      Centro de Investigacion Biomedica en Red de Salud Mental (CIBERSAM), 28029, 
      Madrid, Spain.
FAU - Barrio, R J
AU  - Barrio RJ
AD  - Department of Analytical Chemistry, Faculty of Pharmacy, University of the Basque 
      Country (UPV/EHU), 01006, Vitoria-Gasteiz, Spain.
LA  - eng
PT  - Journal Article
DEP - 20211119
PL  - Netherlands
TA  - Anal Chim Acta
JT  - Analytica chimica acta
JID - 0370534
RN  - 0 (Antibodies)
RN  - 0 (Polymers)
RN  - EC 3.4.21.- (Lactoferrin)
SB  - IM
MH  - Antibodies
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Humans
MH  - Lactoferrin
MH  - *Molecular Imprinting
MH  - *Polymers
OTO - NOTNLM
OT  - ELISA mimic
OT  - Lactoferrin
OT  - Linear polymer
OT  - Plastic antibody
OT  - Reversible deactivation radical polymerisation
OT  - Urine sample
COIS- Declaration of competing interest The authors declare that they have no known 
      competing financial interests or personal relationships that could have appeared 
      to influence the work reported in this paper.
EDAT- 2022/01/17 06:00
MHDA- 2022/01/19 06:00
CRDT- 2022/01/16 20:37
PHST- 2021/07/21 00:00 [received]
PHST- 2021/11/04 00:00 [revised]
PHST- 2021/11/18 00:00 [accepted]
PHST- 2022/01/16 20:37 [entrez]
PHST- 2022/01/17 06:00 [pubmed]
PHST- 2022/01/19 06:00 [medline]
AID - S0003-2670(21)01135-1 [pii]
AID - 10.1016/j.aca.2021.339309 [doi]
PST - ppublish
SO  - Anal Chim Acta. 2022 Jan 25;1191:339309. doi: 10.1016/j.aca.2021.339309. Epub 
      2021 Nov 19.