Cellulose based bio-interfaces for immunodiagnostic applications

Hannes Orelma

Cellulose based bio-interfaces for immunodiagnostic applications

Julkaisun otsikon käännös: Selluloosapohjaiset biointerfaasit immunodiagnostisiin aplikaatioihin

Hannes Orelma

Tutkimustuotos: Doctoral Thesis › Collection of Articles

Abstrakti

Työssä tutkittiin proteiinien vuorovaikutusta pintamuokattujen selluloosamateriaalien kanssa hyödynnettäväksi immunodiagnostisissa sovelluksissa. Erityisesti työssä tutkittiin immunologisten vasta-aineiden kiinnittämistä pysyvästi selluloosapintoihin, ja kiinnitettyjen vasta-aineiden aktiivisuutta tunnistaa antigeenejä, vasta-aineille spesifisiä proteiineja. Edellä mainittujen proteiinien adsorptiota selluloosamateriaaleihin tutkittiin käyttäen pintaherkkiä menetelmiä, kuten kvartsikidevaakaa (QCM-D) ja pintaplasmoniresonanssi-instrumenttia (SPR) selluloosamallipintojen kanssa. Tutkitut proteiinit sitoutuivat modifioimattomiin selluloosapintoihin parhaiten niiden isoelektrisissä pisteissä, mikä osoittaa adsorption tapahtuvan pääasiassa muiden kuin sähköisten vuorovaikutusvoimien välityksellä. Selluloosapinnan kasvanut varaus kasvatti kaikkien tutkittujen proteiinien adsorptiota modifioiduille selluloosapinnoille. Tutkimuksessa selvitettiin myös yhden glykoproteiinin, avidiinin, adsorptiota muokatuille selluloosapinnoille, sekä avidiinin proteiinikuoren oligosakkaridiketjujen vaikutusta adsorptiomekanismeihin. Avidinin adsorptio selluloosapinnalla tapahtui sekä sähköisten ja ei-sähköisten vuorovaikutusten välityksellä ja sitoutuminen oli pysyvää. Tutkimuksessa havaittiin myös oligosakkaridiketjujen vähentävän avidinin adsorptiota selluloosapinnalle. Työssä kehitettiin menetelmiä sitoa vasta-aineita selluloosapintoihin vesifaasissa hyödyntäen ionisia polysakkarideja (karboksymetyyliselluloosa ja kitosaani), jotka adsorboituvat irreversiibelisti selluloosapintoihin. Tutkimuksissa havaittiin, että adsorboimalla tutkittuja ionisia polysakkarideja selluloosapintoihin, voidaan vasta-aineita sitoa selluloosapintaan kovalenttisten sidosten välityksellä. Kehitetyillä biointerfaasilla pystyttiin detektoimaan antigeenejä spesifisesti. Vasta-aineen konformaatio kiinteällä pinnalla vaikuttaa sen kykyyn sitoa tunnistettavaa antigeeniä. Perinteisessä kovalenttisessa immobilisaatiossa vasta-aineen konformaatiota ei pystytä hallitsemaan. Työssä tutkittiin menetelmää kiinnittää vasta-aineita selluloosapintoihin avidini-biotiinisidoksen avulla. Avidini-biotiinisidoksen avulla biointerfaasin kyky tunnistaa antigeenejä saatiin kaksinkertaistettua. Tutkimuksen viimeisessä vaiheessa kehitettiin biointerfaasi hyödyntäen nanoselluloosafilmejä, joiden pinnat modifioitiin käyttäen TEMPO-hapetusta ja EDC/NHS-aktivointia. Tutkitun aktivoidun nanoselluloosakalvon havaittiin sitovan vasta-aineita kovalenttisesti, ja niiden kiinnitys demonstroitiin mustesuihkutulostusta hyödyntäen. Kehitetty biointefaasi tarjoaa lujan ja kestävän alustan tulevaisuuden immunodiagnostisille sovelluksille.

Julkaisun otsikon käännös	Selluloosapohjaiset biointerfaasit immunodiagnostisiin aplikaatioihin
Alkuperäiskieli	Englanti
Pätevyys	Tohtorintutkinto
Myöntävä instituutio	Aalto-yliopisto
Valvoja/neuvonantaja	Laine, Janne, Vastuuprofessori Filpponen, Ilari, Ohjaaja, Ulkoinen henkilö
Kustantaja	Aalto University
Painoksen ISBN	978-952-60-4835-2
Sähköinen ISBN	978-952-60-4836-9
Tila	Julkaistu - 2012
OKM-julkaisutyyppi	G5 Artikkeliväitöskirja

Tutkimusalat

selluloosa
vasta-aine
immobilisointi
immunodiagnostiikka

Pääsy asiakirjaan

http://urn.fi/URN:ISBN:978-952-60-4836-9

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title = "Cellulose based bio-interfaces for immunodiagnostic applications",

abstract = "In this work, the interactions between various proteins and modified cellulose surfaces were investigated. The work focused on the development of immobilization methods for the covalent attachment of specific immunological antibodies (proteins) onto cellulose substrate. The immobilization methods were explored using cellulose model surfaces and surface sensitive techniques, such as quartz crystal microbalance with dissipation monitoring (QCM-D) and surface plasmon resonance (SPR). The highest adsorption of globular proteins on unmodified cellulose surfaces occurred at their respective isoelectric points, suggesting a non-electrostatic adsorption mechanism. An increased surface charge at the cellulose substrate was found to enhance the adsorption of all the proteins investigated. This indicated the presence of attractive electrostatic interactions and the adsorption was found to be mainly irreversible. In addition, the effect of oligosaccharide regions of proteins on their adsorption on cellulose was examined with one glycoprotein, avidin. The adsorption of avidin on cellulose was driven by a combination of electrostatic and non-electrostatic forces, and the adsorption was mainly irreversible. Moreover, the oligosaccharide regions of avidin decreased its adsorption strength to cellulose. In this work, several strategies for covalent immobilization of antibodies onto functionalized cellulose matrices were developed. The novel biointerfaces were capable of sensing antigens both selectively and quantitatively. The use of traditional conjugation chemistries typically leads to a random conformation of immobilized antibodies on the surfaces which in turn may decrease the ability of immobilized antibodies to bind antigens due to the sterical hindrances. Therefore, in this work, the antibodies were immobilized onto cellulose in more oriented manner using avidin-biotin linkage. This approach resulted in over two-fold higher antigen response when compared to those of the traditional conjugation chemistry. In the last part of this work, a biointerface was prepared on a water-resistant nanofibrillar cellulose (NFC) film. The NFC film was made amine reactive by using sequential TEMPO-mediated oxidation and EDC/NHS activation. Activated NFC-films were observed to bind antibodies covalently, and the antibodies could be deposited using standard inkjet printing techniques. The developed NFC-based biointerfaces are expected to open new venues for using cellulose in immunodiagnostic applications.",

keywords = "cellulose, antibodies, immobilization, immunodiagnostic, selluloosa, vasta-aine, immobilisointi, immunodiagnostiikka, cellulose, antibodies, immobilization, immunodiagnostic",

author = "Hannes Orelma",

year = "2012",

language = "English",

isbn = "978-952-60-4835-2",

series = "Aalto University publication series DOCTORAL DISSERTATIONS",

publisher = "Aalto University",

number = "138",

address = "Finland",

school = "Aalto University",

}

TY - BOOK

T1 - Cellulose based bio-interfaces for immunodiagnostic applications

AU - Orelma, Hannes

PY - 2012

Y1 - 2012

N2 - In this work, the interactions between various proteins and modified cellulose surfaces were investigated. The work focused on the development of immobilization methods for the covalent attachment of specific immunological antibodies (proteins) onto cellulose substrate. The immobilization methods were explored using cellulose model surfaces and surface sensitive techniques, such as quartz crystal microbalance with dissipation monitoring (QCM-D) and surface plasmon resonance (SPR). The highest adsorption of globular proteins on unmodified cellulose surfaces occurred at their respective isoelectric points, suggesting a non-electrostatic adsorption mechanism. An increased surface charge at the cellulose substrate was found to enhance the adsorption of all the proteins investigated. This indicated the presence of attractive electrostatic interactions and the adsorption was found to be mainly irreversible. In addition, the effect of oligosaccharide regions of proteins on their adsorption on cellulose was examined with one glycoprotein, avidin. The adsorption of avidin on cellulose was driven by a combination of electrostatic and non-electrostatic forces, and the adsorption was mainly irreversible. Moreover, the oligosaccharide regions of avidin decreased its adsorption strength to cellulose. In this work, several strategies for covalent immobilization of antibodies onto functionalized cellulose matrices were developed. The novel biointerfaces were capable of sensing antigens both selectively and quantitatively. The use of traditional conjugation chemistries typically leads to a random conformation of immobilized antibodies on the surfaces which in turn may decrease the ability of immobilized antibodies to bind antigens due to the sterical hindrances. Therefore, in this work, the antibodies were immobilized onto cellulose in more oriented manner using avidin-biotin linkage. This approach resulted in over two-fold higher antigen response when compared to those of the traditional conjugation chemistry. In the last part of this work, a biointerface was prepared on a water-resistant nanofibrillar cellulose (NFC) film. The NFC film was made amine reactive by using sequential TEMPO-mediated oxidation and EDC/NHS activation. Activated NFC-films were observed to bind antibodies covalently, and the antibodies could be deposited using standard inkjet printing techniques. The developed NFC-based biointerfaces are expected to open new venues for using cellulose in immunodiagnostic applications.

AB - In this work, the interactions between various proteins and modified cellulose surfaces were investigated. The work focused on the development of immobilization methods for the covalent attachment of specific immunological antibodies (proteins) onto cellulose substrate. The immobilization methods were explored using cellulose model surfaces and surface sensitive techniques, such as quartz crystal microbalance with dissipation monitoring (QCM-D) and surface plasmon resonance (SPR). The highest adsorption of globular proteins on unmodified cellulose surfaces occurred at their respective isoelectric points, suggesting a non-electrostatic adsorption mechanism. An increased surface charge at the cellulose substrate was found to enhance the adsorption of all the proteins investigated. This indicated the presence of attractive electrostatic interactions and the adsorption was found to be mainly irreversible. In addition, the effect of oligosaccharide regions of proteins on their adsorption on cellulose was examined with one glycoprotein, avidin. The adsorption of avidin on cellulose was driven by a combination of electrostatic and non-electrostatic forces, and the adsorption was mainly irreversible. Moreover, the oligosaccharide regions of avidin decreased its adsorption strength to cellulose. In this work, several strategies for covalent immobilization of antibodies onto functionalized cellulose matrices were developed. The novel biointerfaces were capable of sensing antigens both selectively and quantitatively. The use of traditional conjugation chemistries typically leads to a random conformation of immobilized antibodies on the surfaces which in turn may decrease the ability of immobilized antibodies to bind antigens due to the sterical hindrances. Therefore, in this work, the antibodies were immobilized onto cellulose in more oriented manner using avidin-biotin linkage. This approach resulted in over two-fold higher antigen response when compared to those of the traditional conjugation chemistry. In the last part of this work, a biointerface was prepared on a water-resistant nanofibrillar cellulose (NFC) film. The NFC film was made amine reactive by using sequential TEMPO-mediated oxidation and EDC/NHS activation. Activated NFC-films were observed to bind antibodies covalently, and the antibodies could be deposited using standard inkjet printing techniques. The developed NFC-based biointerfaces are expected to open new venues for using cellulose in immunodiagnostic applications.

KW - cellulose

KW - antibodies

KW - immobilization

KW - immunodiagnostic

KW - selluloosa

KW - vasta-aine

KW - immobilisointi

KW - immunodiagnostiikka

KW - cellulose

KW - antibodies

KW - immobilization

KW - immunodiagnostic

M3 - Doctoral Thesis

SN - 978-952-60-4835-2

T3 - Aalto University publication series DOCTORAL DISSERTATIONS

PB - Aalto University

ER -

Cellulose based bio-interfaces for immunodiagnostic applications

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