KFU. Card publication. Inhibition of nonspecific polymerase activity using Poly(Aspartic) acid as a model anionic polyelectrolyte

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FEDERAL UNIVERSITY

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INHIBITION OF NONSPECIFIC POLYMERASE ACTIVITY USING POLY(ASPARTIC) ACID AS A MODEL ANIONIC POLYELECTROLYTE

Form of presentation

Articles in international journals and collections

Year of publication

2021

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Authors, employees of KFU

Kamalov Marat Ilyasovich, author

Bibliographic description in the original language

Assol R. Sakhabutdinova, Marat I. Kamalov, Diana V. Salakhieva, Ayrat R. Mavzyutov, Ravil R. Garafutdinov, Inhibition of nonspecific polymerase activity using Poly(Aspartic) acid as a model anionic polyelectrolyte, Analytical Biochemistry, Volume 628, 2021, 114267, ISSN 0003-2697,https://doi.org/10.1016/j.ab.2021.114267.

Annotation

DNA polymerases with strand-displacement activity allow to amplify nucleic acids under isothermal conditions but often lead to undesirable by-products. Here, we report the increase of specificity of isothermal amplification in the presence of poly (aspartic) acids (pAsp). We hypothesized that side reactions occur due to the binding of the phosphate backbone of synthesized DNA strands with surface amino groups of the polymerase, and weakly acidic polyelectrolytes could shield polymerase molecules from DNA and thereby inhibit nonspecific amplification. Suppression of nonspecific polymerase activity by pAsp was studied on multimerization as a model side reaction. It was found that a low concentration of pAsp (0.01%) provides successful amplification of specific DNA targets. The inhibitory effect of pAsp is due to its polymeric structure since aspartic acid did affect neither specific nor nonspecific amplification. Strongly acidic polyelectrolyte heparin does not possess the same selectivity since it suppresses any DNA synthesis. The applicability of pAsp to prevent nonspecific reactions and reliable detection of the specific target has been demonstrated on the genetic material of SARS-CoV-2 coronavirus using Loop-mediated isothermal amplification.

Keywords

Isothermal amplification, specificity, multimerization, poly(aspartic) acid, coronavirus SARS-CoV-2

The name of the journal

Analytical Biochemistry

URL

https://www.scopus.com/inward/record.uri?eid=2-s2.0-85108239605&doi=10.1016%2fj.ab.2021.114267&partnerID=40&md5=729b1903102dc4060b8e1351a1a6018c

Please use this ID to quote from or refer to the card

https://repository.kpfu.ru/eng/?p_id=283247&p_lang=2

Full metadata record

Field DC	Value	Language
dc.contributor.author	Kamalov Marat Ilyasovich	ru_RU
dc.date.accessioned	2021-01-01T00:00:00Z	ru_RU
dc.date.available	2021-01-01T00:00:00Z	ru_RU
dc.date.issued	2021	ru_RU
dc.identifier.citation	Assol R. Sakhabutdinova, Marat I. Kamalov, Diana V. Salakhieva, Ayrat R. Mavzyutov, Ravil R. Garafutdinov, Inhibition of nonspecific polymerase activity using Poly(Aspartic) acid as a model anionic polyelectrolyte, Analytical Biochemistry, Volume 628, 2021, 114267, ISSN 0003-2697,https://doi.org/10.1016/j.ab.2021.114267.	ru_RU
dc.identifier.uri	https://repository.kpfu.ru/eng/?p_id=283247&p_lang=2	ru_RU
dc.description.abstract	Analytical Biochemistry	ru_RU
dc.description.abstract	DNA polymerases with strand-displacement activity allow to amplify nucleic acids under isothermal conditions but often lead to undesirable by-products. Here, we report the increase of specificity of isothermal amplification in the presence of poly (aspartic) acids (pAsp). We hypothesized that side reactions occur due to the binding of the phosphate backbone of synthesized DNA strands with surface amino groups of the polymerase, and weakly acidic polyelectrolytes could shield polymerase molecules from DNA and thereby inhibit nonspecific amplification. Suppression of nonspecific polymerase activity by pAsp was studied on multimerization as a model side reaction. It was found that a low concentration of pAsp (0.01%) provides successful amplification of specific DNA targets. The inhibitory effect of pAsp is due to its polymeric structure since aspartic acid did affect neither specific nor nonspecific amplification. Strongly acidic polyelectrolyte heparin does not possess the same selectivity since it suppresses any DNA synthesis. The applicability of pAsp to prevent nonspecific reactions and reliable detection of the specific target has been demonstrated on the genetic material of SARS-CoV-2 coronavirus using Loop-mediated isothermal amplification.	ru_RU
dc.language.iso	ru	ru_RU
dc.subject	Isothermal amplification	ru_RU
dc.subject	specificity	ru_RU
dc.subject	multimerization	ru_RU
dc.subject	poly(aspartic) acid	ru_RU
dc.subject	coronavirus SARS-CoV-2	ru_RU
dc.title	Inhibition of nonspecific polymerase activity using Poly(Aspartic) acid as a model anionic polyelectrolyte	ru_RU
dc.type	Articles in international journals and collections	ru_RU