Novel binders derived from an albumin-binding domain scaffold targeting human prostate secretory protein 94 (PSP94)

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F86652036%3A_____%2F15%3A00464077" target="_blank" >RIV/86652036:_____/15:00464077 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/61388971:_____/15:00464077

Výsledek na webu

<a href="http://dx.doi.org/10.1007/s13238-015-0194-9" target="_blank" >http://dx.doi.org/10.1007/s13238-015-0194-9</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1007/s13238-015-0194-9" target="_blank" >10.1007/s13238-015-0194-9</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Novel binders derived from an albumin-binding domain scaffold targeting human prostate secretory protein 94 (PSP94)

Popis výsledku v původním jazyce

Prostate secretory protein 94 (PSP94), a dominant protein of human seminal plasma known also as β-microseminoprotein (MSMB), has recently been shown to decrease serum level expression during prostate cancer (PC) progression and indicated as a novel promising serum oncomarker with a predictive role in PC diagnosis. In this work we aimed to generate a novel class of protein binders targeted to human PSP94 that can serve as robust capture proteins for improved PC diagnostics. We used high-complex combinatorial library derived from an albumin-binding domain (ABD) scaffold of streptococcal protein G with a theoretical complexity up to 2 x 1014 protein variants and ribosome display to generate a plasmid library of selected variants. Individual clones were characterized by binding of ABD-derived proteins to recombinant PSP94 and sequentially compared and characterized. Collection of 35 ABD-derived protein binders of human PSP94 (called PAB binders), corresponding to 29 different sequence variants, was generated. PAB036, PAB046 and PAB050 were identified as the most promising binding candidates characterized in ELISA. Their stability was verified by thermal shift assay and the binding affinity to the recombinant PSP94 was estimated by microscale thermophoresis. These PAB variants bound to PSP94-expressing human LNCaP prostate carcinoma cells and this binding was dose-dependently inhibited by recombinant PSP94. Thus, we generated a unique collection of novel proteins binders of human PSP94 that can serve as a valuable tool for human PSP94 detection and, with a further modification, as high-affinity capture proteins for the development of biosensors for more complex PC diagnostics.

Název v anglickém jazyce

Novel binders derived from an albumin-binding domain scaffold targeting human prostate secretory protein 94 (PSP94)

Popis výsledku anglicky

Prostate secretory protein 94 (PSP94), a dominant protein of human seminal plasma known also as β-microseminoprotein (MSMB), has recently been shown to decrease serum level expression during prostate cancer (PC) progression and indicated as a novel promising serum oncomarker with a predictive role in PC diagnosis. In this work we aimed to generate a novel class of protein binders targeted to human PSP94 that can serve as robust capture proteins for improved PC diagnostics. We used high-complex combinatorial library derived from an albumin-binding domain (ABD) scaffold of streptococcal protein G with a theoretical complexity up to 2 x 1014 protein variants and ribosome display to generate a plasmid library of selected variants. Individual clones were characterized by binding of ABD-derived proteins to recombinant PSP94 and sequentially compared and characterized. Collection of 35 ABD-derived protein binders of human PSP94 (called PAB binders), corresponding to 29 different sequence variants, was generated. PAB036, PAB046 and PAB050 were identified as the most promising binding candidates characterized in ELISA. Their stability was verified by thermal shift assay and the binding affinity to the recombinant PSP94 was estimated by microscale thermophoresis. These PAB variants bound to PSP94-expressing human LNCaP prostate carcinoma cells and this binding was dose-dependently inhibited by recombinant PSP94. Thus, we generated a unique collection of novel proteins binders of human PSP94 that can serve as a valuable tool for human PSP94 detection and, with a further modification, as high-affinity capture proteins for the development of biosensors for more complex PC diagnostics.

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

EB - Genetika a molekulární biologie

OECD FORD obor

—

Projekt

—

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2015

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Protein & Cell

ISSN

1674-800X

e-ISSN

—

Svazek periodika

6

Číslo periodika v rámci svazku

10

Stát vydavatele periodika

CN - Čínská lidová republika

Počet stran výsledku

6

Strana od-do

774-779

Kód UT WoS článku

000362653100010

EID výsledku v databázi Scopus

—