A NEW METHOD BASED ON THE USE OF IMMOBILIZED SINGLE-DOMAIN ANTIBODIES TO REMOVE CERTAIN MAJOR PROTEINS FROM BLOOD PLASMA HELPS TO REDUCE NONSPECIFIC SIGNAL IN AN IMMUNOASSAY

A generally underestimated problem of immunoassays is its susceptibility to various interferences, being the Achilles heel of these assays (interventions in the analysis). The presence of interfering substances in the patient’s specimen can cause erroneous test sample result, which may lead to incor...

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Autores principales:	O. S. Goryainova, E. O. Khan, T. I. Ivanova, S. V. Tillib
Formato:	article
Lenguaje:	RU
Publicado:	SPb RAACI 2019
Materias:	single-domain antibodies immunosorbents blood plasma pre-treatment immunoassay Immunologic diseases. Allergy RC581-607
Acceso en línea:	https://doaj.org/article/2f38676583af488fb70b47952d495143
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spelling	oai:doaj.org-article:2f38676583af488fb70b47952d4951432021-11-18T08:03:48ZA NEW METHOD BASED ON THE USE OF IMMOBILIZED SINGLE-DOMAIN ANTIBODIES TO REMOVE CERTAIN MAJOR PROTEINS FROM BLOOD PLASMA HELPS TO REDUCE NONSPECIFIC SIGNAL IN AN IMMUNOASSAY1563-06252313-741X10.15789/1563-0625-2019-3-567-575https://doaj.org/article/2f38676583af488fb70b47952d4951432019-07-01T00:00:00Zhttps://www.mimmun.ru/mimmun/article/view/1831https://doaj.org/toc/1563-0625https://doaj.org/toc/2313-741XA generally underestimated problem of immunoassays is its susceptibility to various interferences, being the Achilles heel of these assays (interventions in the analysis). The presence of interfering substances in the patient’s specimen can cause erroneous test sample result, which may lead to incorrect diagnosis and catastrophic consequences for the patient. Hence, one should pay particular attention to identifying possible interferences in the test systems used and, when possible, develop and apply methods to overcome them. The issue of avoiding interference is particularly important for immunoassays of biomarkers in human plasma or serum. In order to reduce possible interferences, it would be desirable to have an opportunity of specific and adaptable pretreatment of blood samples for a specifically assayed marker protein, as applied to a specific test system. We assume that such pretreatment may be done by combining the immunosorbents based on the use of special single-domain antibodies (nanobodies).The nanobodies are recombinant proteins, derivatives of single-domain antigen-recognizing variable fragments of specific antibodies, consisting of a dimer of truncated heavy chains in the complete absence of light chains. Such specific antibodies are detectable in the normal samples taken from members of the Camelidae family (Camelids), and in some species of cartilaginous fishes, along with the common antibodies. The special properties of nanobodies can provide certain advantages in their use, compared with antibodies of traditional structure and their derivatives.In this paper, we have shown for the first time, that the immunosorbents based on certain combination of single-domain antibodies used as ligands able for specifically binding and removal of specific high-abundance human blood proteins, may be selected for a given marker blood antigen in such a way that they will be an effective tool for blood pretreatment, aiming to reduce possible effects of interference and increased sensitivity in the diagnostic “sandwich” enzyme immunoassay. A new method of plasma pretreatment is demonstrated with human plasma samples (at a dilution of 1:40) of two patients. A previously developed model system for detection of lactoferrin protein was used to analyze plasma samples. It is shown that a significantly increased ratio of total detected signal to the nonspecific background signal could be obtained after drastic reduction of this background by affinity removal of 3 major protein fractions, i.e., fibrinogen, IgG alpha-2-macroglobulin from blood plasma samples, using appropriate immobilized single domain antibodies.O. S. GoryainovaE. O. KhanT. I. IvanovaS. V. TillibSPb RAACIarticlesingle-domain antibodiesimmunosorbentsblood plasma pre-treatmentimmunoassayImmunologic diseases. AllergyRC581-607RUMedicinskaâ Immunologiâ, Vol 21, Iss 3, Pp 567-575 (2019)
institution	DOAJ
collection	DOAJ
language	RU
topic	single-domain antibodies immunosorbents blood plasma pre-treatment immunoassay Immunologic diseases. Allergy RC581-607
spellingShingle	single-domain antibodies immunosorbents blood plasma pre-treatment immunoassay Immunologic diseases. Allergy RC581-607 O. S. Goryainova E. O. Khan T. I. Ivanova S. V. Tillib A NEW METHOD BASED ON THE USE OF IMMOBILIZED SINGLE-DOMAIN ANTIBODIES TO REMOVE CERTAIN MAJOR PROTEINS FROM BLOOD PLASMA HELPS TO REDUCE NONSPECIFIC SIGNAL IN AN IMMUNOASSAY
description	A generally underestimated problem of immunoassays is its susceptibility to various interferences, being the Achilles heel of these assays (interventions in the analysis). The presence of interfering substances in the patient’s specimen can cause erroneous test sample result, which may lead to incorrect diagnosis and catastrophic consequences for the patient. Hence, one should pay particular attention to identifying possible interferences in the test systems used and, when possible, develop and apply methods to overcome them. The issue of avoiding interference is particularly important for immunoassays of biomarkers in human plasma or serum. In order to reduce possible interferences, it would be desirable to have an opportunity of specific and adaptable pretreatment of blood samples for a specifically assayed marker protein, as applied to a specific test system. We assume that such pretreatment may be done by combining the immunosorbents based on the use of special single-domain antibodies (nanobodies).The nanobodies are recombinant proteins, derivatives of single-domain antigen-recognizing variable fragments of specific antibodies, consisting of a dimer of truncated heavy chains in the complete absence of light chains. Such specific antibodies are detectable in the normal samples taken from members of the Camelidae family (Camelids), and in some species of cartilaginous fishes, along with the common antibodies. The special properties of nanobodies can provide certain advantages in their use, compared with antibodies of traditional structure and their derivatives.In this paper, we have shown for the first time, that the immunosorbents based on certain combination of single-domain antibodies used as ligands able for specifically binding and removal of specific high-abundance human blood proteins, may be selected for a given marker blood antigen in such a way that they will be an effective tool for blood pretreatment, aiming to reduce possible effects of interference and increased sensitivity in the diagnostic “sandwich” enzyme immunoassay. A new method of plasma pretreatment is demonstrated with human plasma samples (at a dilution of 1:40) of two patients. A previously developed model system for detection of lactoferrin protein was used to analyze plasma samples. It is shown that a significantly increased ratio of total detected signal to the nonspecific background signal could be obtained after drastic reduction of this background by affinity removal of 3 major protein fractions, i.e., fibrinogen, IgG alpha-2-macroglobulin from blood plasma samples, using appropriate immobilized single domain antibodies.
format	article
author	O. S. Goryainova E. O. Khan T. I. Ivanova S. V. Tillib
author_facet	O. S. Goryainova E. O. Khan T. I. Ivanova S. V. Tillib
author_sort	O. S. Goryainova
title	A NEW METHOD BASED ON THE USE OF IMMOBILIZED SINGLE-DOMAIN ANTIBODIES TO REMOVE CERTAIN MAJOR PROTEINS FROM BLOOD PLASMA HELPS TO REDUCE NONSPECIFIC SIGNAL IN AN IMMUNOASSAY
title_short	A NEW METHOD BASED ON THE USE OF IMMOBILIZED SINGLE-DOMAIN ANTIBODIES TO REMOVE CERTAIN MAJOR PROTEINS FROM BLOOD PLASMA HELPS TO REDUCE NONSPECIFIC SIGNAL IN AN IMMUNOASSAY
title_full	A NEW METHOD BASED ON THE USE OF IMMOBILIZED SINGLE-DOMAIN ANTIBODIES TO REMOVE CERTAIN MAJOR PROTEINS FROM BLOOD PLASMA HELPS TO REDUCE NONSPECIFIC SIGNAL IN AN IMMUNOASSAY
title_fullStr	A NEW METHOD BASED ON THE USE OF IMMOBILIZED SINGLE-DOMAIN ANTIBODIES TO REMOVE CERTAIN MAJOR PROTEINS FROM BLOOD PLASMA HELPS TO REDUCE NONSPECIFIC SIGNAL IN AN IMMUNOASSAY
title_full_unstemmed	A NEW METHOD BASED ON THE USE OF IMMOBILIZED SINGLE-DOMAIN ANTIBODIES TO REMOVE CERTAIN MAJOR PROTEINS FROM BLOOD PLASMA HELPS TO REDUCE NONSPECIFIC SIGNAL IN AN IMMUNOASSAY
title_sort	new method based on the use of immobilized single-domain antibodies to remove certain major proteins from blood plasma helps to reduce nonspecific signal in an immunoassay
publisher	SPb RAACI
publishDate	2019
url	https://doaj.org/article/2f38676583af488fb70b47952d495143
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A NEW METHOD BASED ON THE USE OF IMMOBILIZED SINGLE-DOMAIN ANTIBODIES TO REMOVE CERTAIN MAJOR PROTEINS FROM BLOOD PLASMA HELPS TO REDUCE NONSPECIFIC SIGNAL IN AN IMMUNOASSAY

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