Purification and in vitro evaluation of an anti-HER2 affibody-monomethyl auristatin E conjugate in HER2-positive cancer cells

2021
journal article
article
5
cris.lastimport.wos2024-04-10T02:01:05Z
dc.abstract.enA promising approach for the development of high-affinity tumor targeting ADCs is the use of engineered protein drugs, such as affibody molecules, which represent a valuable alternative to monoclonal antibodies (mAbs) in cancer-targeted therapy. We developed a method for a more efficient purification of the $Z_{HER2:2891}DCS$ affibody conjugated with the cytotoxic antimitotic agent auristatin E (MMAE), and its efficacy was tested in vitro on cell viability, proliferation, migration, and apoptosis. The effects of $Z_{HER2:2891}DCS$-MMAE were compared with the clinically approved monoclonal antibody trastuzumab ($Herceptin^{®}$). To demonstrate that $Z_{HER2:2891}DCS$-MMAE can selectively target HER2 overexpressing tumor cells, we used three different cell lines: the human adenocarcinoma cell lines SK-BR-3 and ZR-75-1, both overexpressing HER2, and the triple-negative breast cancer cell line MDA-MB-231. MTT assay showed that $Z_{HER2:2891}DCS$-MMAE induces a significant time-dependent toxic effect in SK-BR-3 cells. A 30% reduction of cell viability was already found after 10 min exposure at a concentration of 7 nM ($IC_{50}$ of 80.2 nM). On the contrary, MDA-MB-231 cells, which express basal levels of HER2, were not affected by the conjugate. The cytotoxic effect of the $Z_{HER2:2891}DCS$-MMAE was confirmed by measuring apoptosis by flow cytometry. In SK-BR-3 cells, increasing concentrations of conjugated affibody induced cell death starting from 10 min of treatment, with the strongest effect observed after 48 h. Overall, these results demonstrate that the ADC, formed by the anti-HER2 affibody conjugated to monomethyl auristatin E, efficiently interacts with high affinity with HER2 positive cancer cells in vitro, allowing the selective and specific delivery of the cytotoxic payload.pl
dc.affiliationWydział Biochemii, Biofizyki i Biotechnologii : Zakład Mikrobiologiipl
dc.contributor.authorDamiani, Isabellapl
dc.contributor.authorCastiglioni, Silviapl
dc.contributor.authorSochaj-Gregorczyk, Alicja - 377819 pl
dc.contributor.authorBonacina, Fabriziapl
dc.contributor.authorColombo, Irmapl
dc.contributor.authorRusconi, Valentinapl
dc.contributor.authorOtlewski, Jacekpl
dc.contributor.authorCorsini, Albertopl
dc.contributor.authorBellosta, Stefanopl
dc.date.accessioned2021-10-29T12:43:09Z
dc.date.available2021-10-29T12:43:09Z
dc.date.issued2021pl
dc.date.openaccess0
dc.description.accesstimew momencie opublikowania
dc.description.number8pl
dc.description.versionostateczna wersja wydawcy
dc.description.volume10pl
dc.identifier.articleid758pl
dc.identifier.doi10.3390/biology10080758pl
dc.identifier.eissn2079-7737pl
dc.identifier.projectROD UJ / Opl
dc.identifier.urihttps://ruj.uj.edu.pl/xmlui/handle/item/282284
dc.languageengpl
dc.language.containerengpl
dc.rightsUdzielam licencji. Uznanie autorstwa 4.0 Międzynarodowa*
dc.rights.licenceCC-BY
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/legalcode.pl*
dc.share.typeotwarte czasopismo
dc.subject.enaffibodypl
dc.subject.enbreast cancerpl
dc.subject.enHER2 overexpressionpl
dc.subject.entarget therapypl
dc.subtypeArticlepl
dc.titlePurification and in vitro evaluation of an anti-HER2 affibody-monomethyl auristatin E conjugate in HER2-positive cancer cellspl
dc.title.journalBiologypl
dc.typeJournalArticlepl
dspace.entity.typePublication
cris.lastimport.wos
2024-04-10T02:01:05Z
dc.abstract.enpl
A promising approach for the development of high-affinity tumor targeting ADCs is the use of engineered protein drugs, such as affibody molecules, which represent a valuable alternative to monoclonal antibodies (mAbs) in cancer-targeted therapy. We developed a method for a more efficient purification of the $Z_{HER2:2891}DCS$ affibody conjugated with the cytotoxic antimitotic agent auristatin E (MMAE), and its efficacy was tested in vitro on cell viability, proliferation, migration, and apoptosis. The effects of $Z_{HER2:2891}DCS$-MMAE were compared with the clinically approved monoclonal antibody trastuzumab ($Herceptin^{®}$). To demonstrate that $Z_{HER2:2891}DCS$-MMAE can selectively target HER2 overexpressing tumor cells, we used three different cell lines: the human adenocarcinoma cell lines SK-BR-3 and ZR-75-1, both overexpressing HER2, and the triple-negative breast cancer cell line MDA-MB-231. MTT assay showed that $Z_{HER2:2891}DCS$-MMAE induces a significant time-dependent toxic effect in SK-BR-3 cells. A 30% reduction of cell viability was already found after 10 min exposure at a concentration of 7 nM ($IC_{50}$ of 80.2 nM). On the contrary, MDA-MB-231 cells, which express basal levels of HER2, were not affected by the conjugate. The cytotoxic effect of the $Z_{HER2:2891}DCS$-MMAE was confirmed by measuring apoptosis by flow cytometry. In SK-BR-3 cells, increasing concentrations of conjugated affibody induced cell death starting from 10 min of treatment, with the strongest effect observed after 48 h. Overall, these results demonstrate that the ADC, formed by the anti-HER2 affibody conjugated to monomethyl auristatin E, efficiently interacts with high affinity with HER2 positive cancer cells in vitro, allowing the selective and specific delivery of the cytotoxic payload.
dc.affiliationpl
Wydział Biochemii, Biofizyki i Biotechnologii : Zakład Mikrobiologii
dc.contributor.authorpl
Damiani, Isabella
dc.contributor.authorpl
Castiglioni, Silvia
dc.contributor.authorpl
Sochaj-Gregorczyk, Alicja - 377819
dc.contributor.authorpl
Bonacina, Fabrizia
dc.contributor.authorpl
Colombo, Irma
dc.contributor.authorpl
Rusconi, Valentina
dc.contributor.authorpl
Otlewski, Jacek
dc.contributor.authorpl
Corsini, Alberto
dc.contributor.authorpl
Bellosta, Stefano
dc.date.accessioned
2021-10-29T12:43:09Z
dc.date.available
2021-10-29T12:43:09Z
dc.date.issuedpl
2021
dc.date.openaccess
0
dc.description.accesstime
w momencie opublikowania
dc.description.numberpl
8
dc.description.version
ostateczna wersja wydawcy
dc.description.volumepl
10
dc.identifier.articleidpl
758
dc.identifier.doipl
10.3390/biology10080758
dc.identifier.eissnpl
2079-7737
dc.identifier.projectpl
ROD UJ / O
dc.identifier.uri
https://ruj.uj.edu.pl/xmlui/handle/item/282284
dc.languagepl
eng
dc.language.containerpl
eng
dc.rights*
Udzielam licencji. Uznanie autorstwa 4.0 Międzynarodowa
dc.rights.licence
CC-BY
dc.rights.uri*
http://creativecommons.org/licenses/by/4.0/legalcode.pl
dc.share.type
otwarte czasopismo
dc.subject.enpl
affibody
dc.subject.enpl
breast cancer
dc.subject.enpl
HER2 overexpression
dc.subject.enpl
target therapy
dc.subtypepl
Article
dc.titlepl
Purification and in vitro evaluation of an anti-HER2 affibody-monomethyl auristatin E conjugate in HER2-positive cancer cells
dc.title.journalpl
Biology
dc.typepl
JournalArticle
dspace.entity.type
Publication
Affiliations

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