Engineered Fully Human Single-Chain Monoclonal Antibodies to PIM2 Kinase

Proviral integration site of Moloney virus-2 (PIM2) is overexpressed in multiple human cancer cells and high level is related to poor prognosis; thus, PIM2 kinase is a rational target of anti-cancer therapeutics. Several chemical inhibitors targeting PIMs/PIM2 or their downstream signaling molecules...

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Autores principales:	Kanasap Kaewchim, Kittirat Glab-ampai, Kodchakorn Mahasongkram, Monrat Chulanetra, Watee Seesuay, Wanpen Chaicumpa, Nitat Sookrung
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	human scFv phage display PIM2 kinase ATP-binding pocket homology modeling intermolecular docking Organic chemistry QD241-441
Acceso en línea:	https://doaj.org/article/ba7c415806c34e539f9b7885d440a0cf
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spelling	oai:doaj.org-article:ba7c415806c34e539f9b7885d440a0cf2021-11-11T18:27:03ZEngineered Fully Human Single-Chain Monoclonal Antibodies to PIM2 Kinase10.3390/molecules262164361420-3049https://doaj.org/article/ba7c415806c34e539f9b7885d440a0cf2021-10-01T00:00:00Zhttps://www.mdpi.com/1420-3049/26/21/6436https://doaj.org/toc/1420-3049Proviral integration site of Moloney virus-2 (PIM2) is overexpressed in multiple human cancer cells and high level is related to poor prognosis; thus, PIM2 kinase is a rational target of anti-cancer therapeutics. Several chemical inhibitors targeting PIMs/PIM2 or their downstream signaling molecules have been developed for treatment of different cancers. However, their off-target toxicity is common in clinical trials, so they could not be advanced to official approval for clinical application. Here, we produced human single-chain antibody fragments (HuscFvs) to PIM2 by using phage display library, which was constructed in a way that a portion of phages in the library carried HuscFvs against human own proteins on their surface with the respective antibody genes in the phage genome. Bacterial derived-recombinant PIM2 (rPIM2) was used as an antigenic bait to fish out the rPIM2-bound phages from the library. Three <i>E. coli</i> clones transfected with the HuscFv genes derived from the rPIM2-bound phages expressed HuscFvs that bound also to native PIM2 from cancer cells. The HuscFvs presumptively interact with the PIM2 at the ATP binding pocket and kinase active loop. They were as effective as small chemical drug inhibitor (AZD1208, which is an ATP competitive inhibitor of all PIM isoforms for <i>ex vivo</i> use) in inhibiting PIM kinase activity. The HuscFvs should be engineered into a cell-penetrating format and tested further towards clinical application as a novel and safe pan-anti-cancer therapeutics.Kanasap KaewchimKittirat Glab-ampaiKodchakorn MahasongkramMonrat ChulanetraWatee SeesuayWanpen ChaicumpaNitat SookrungMDPI AGarticlehuman scFvphage displayPIM2 kinaseATP-binding pockethomology modelingintermolecular dockingOrganic chemistryQD241-441ENMolecules, Vol 26, Iss 6436, p 6436 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	human scFv phage display PIM2 kinase ATP-binding pocket homology modeling intermolecular docking Organic chemistry QD241-441
spellingShingle	human scFv phage display PIM2 kinase ATP-binding pocket homology modeling intermolecular docking Organic chemistry QD241-441 Kanasap Kaewchim Kittirat Glab-ampai Kodchakorn Mahasongkram Monrat Chulanetra Watee Seesuay Wanpen Chaicumpa Nitat Sookrung Engineered Fully Human Single-Chain Monoclonal Antibodies to PIM2 Kinase
description	Proviral integration site of Moloney virus-2 (PIM2) is overexpressed in multiple human cancer cells and high level is related to poor prognosis; thus, PIM2 kinase is a rational target of anti-cancer therapeutics. Several chemical inhibitors targeting PIMs/PIM2 or their downstream signaling molecules have been developed for treatment of different cancers. However, their off-target toxicity is common in clinical trials, so they could not be advanced to official approval for clinical application. Here, we produced human single-chain antibody fragments (HuscFvs) to PIM2 by using phage display library, which was constructed in a way that a portion of phages in the library carried HuscFvs against human own proteins on their surface with the respective antibody genes in the phage genome. Bacterial derived-recombinant PIM2 (rPIM2) was used as an antigenic bait to fish out the rPIM2-bound phages from the library. Three <i>E. coli</i> clones transfected with the HuscFv genes derived from the rPIM2-bound phages expressed HuscFvs that bound also to native PIM2 from cancer cells. The HuscFvs presumptively interact with the PIM2 at the ATP binding pocket and kinase active loop. They were as effective as small chemical drug inhibitor (AZD1208, which is an ATP competitive inhibitor of all PIM isoforms for <i>ex vivo</i> use) in inhibiting PIM kinase activity. The HuscFvs should be engineered into a cell-penetrating format and tested further towards clinical application as a novel and safe pan-anti-cancer therapeutics.
format	article
author	Kanasap Kaewchim Kittirat Glab-ampai Kodchakorn Mahasongkram Monrat Chulanetra Watee Seesuay Wanpen Chaicumpa Nitat Sookrung
author_facet	Kanasap Kaewchim Kittirat Glab-ampai Kodchakorn Mahasongkram Monrat Chulanetra Watee Seesuay Wanpen Chaicumpa Nitat Sookrung
author_sort	Kanasap Kaewchim
title	Engineered Fully Human Single-Chain Monoclonal Antibodies to PIM2 Kinase
title_short	Engineered Fully Human Single-Chain Monoclonal Antibodies to PIM2 Kinase
title_full	Engineered Fully Human Single-Chain Monoclonal Antibodies to PIM2 Kinase
title_fullStr	Engineered Fully Human Single-Chain Monoclonal Antibodies to PIM2 Kinase
title_full_unstemmed	Engineered Fully Human Single-Chain Monoclonal Antibodies to PIM2 Kinase
title_sort	engineered fully human single-chain monoclonal antibodies to pim2 kinase
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/ba7c415806c34e539f9b7885d440a0cf
work_keys_str_mv	AT kanasapkaewchim engineeredfullyhumansinglechainmonoclonalantibodiestopim2kinase AT kittiratglabampai engineeredfullyhumansinglechainmonoclonalantibodiestopim2kinase AT kodchakornmahasongkram engineeredfullyhumansinglechainmonoclonalantibodiestopim2kinase AT monratchulanetra engineeredfullyhumansinglechainmonoclonalantibodiestopim2kinase AT wateeseesuay engineeredfullyhumansinglechainmonoclonalantibodiestopim2kinase AT wanpenchaicumpa engineeredfullyhumansinglechainmonoclonalantibodiestopim2kinase AT nitatsookrung engineeredfullyhumansinglechainmonoclonalantibodiestopim2kinase
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Engineered Fully Human Single-Chain Monoclonal Antibodies to PIM2 Kinase

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