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Pregled bibliografske jedinice broj: 812217

Impact of dissociation constant on immunoaffinity and affinity chromatography


Sviben, Dora; Halassy, Beata; Forčić, Dubravko; Brgles, Marija
Impact of dissociation constant on immunoaffinity and affinity chromatography // Symposium & Workshop on Microscale Thermophoresis : book of abstracts / Godinić Mikulčić, Vlatka ; Sviben, Igor ; Rokov Plavec, Jasmina (ur.).
Zagreb: University of Zagreb, Faculty of Science, Chemistry Department, Division of Biochemistry, 2015. str. 31-31 (poster, nije recenziran, sažetak, znanstveni)


CROSBI ID: 812217 Za ispravke kontaktirajte CROSBI podršku putem web obrasca

Naslov
Impact of dissociation constant on immunoaffinity and affinity chromatography

Autori
Sviben, Dora ; Halassy, Beata ; Forčić, Dubravko ; Brgles, Marija

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Symposium & Workshop on Microscale Thermophoresis : book of abstracts / Godinić Mikulčić, Vlatka ; Sviben, Igor ; Rokov Plavec, Jasmina - Zagreb : University of Zagreb, Faculty of Science, Chemistry Department, Division of Biochemistry, 2015, 31-31

ISBN
978-953-6076-37-6

Skup
Symposium and Workshop on Microscale Thermophoresis

Mjesto i datum
Zagreb, Hrvatska, 30.06.2015. - 01.07.2015

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
affinity chromatography ; immunoaffinity chromatography ; dissociation constant

Sažetak
Affinity chromatography is a powerful purification method based on specific interaction of an e.g. enzyme and substrate, receptor and ligand, or antibody and antigen (immunoaffinity chromatography). Advantages of immunoaffinity chromatography are that it can be used for any protein (or even virus) regardless of its function and a very high binding affinity of antigen and antibody enabling isolation of a highly pure interacting partner. Disadvantage of the immunoaffinity chromatography is that a specific competitive elution cannot be employed as is the case with affinity chromatography employing e.g. enzyme or a receptor. Also, high affinity antigen- antibody interactions can only be disrupted using extreme conditions such as low pH conditions, denaturating agents or chaotropic salts. These conditions affect all noncovalent interactions, not only intermolecular but also intramolecular, potentially causing damage to the immobilized protein and the protein to be isolated making immunoaffinity inappropriate for general use. We aimed to find elution conditions disrupting antigen-antibody interactions but under native conditions. For testing of these elution conditions we used two immunoaffinity columns. In both cases columns were prepared by binding of antibodies to epoxy-activated monolith column. One column was specific for ovalbumin and the other for mumps virus. For comparison we also used tested elution systems with commercial protein G column and immunoglobulin solution. Effectiveness of tested elution systems was found different in these three columns, but the trend was the same. This indicates that possibly the differences in dissociation constants i.e. interaction strength are reflected in the obtained results regarding effectiveness of tested elution systems. So we would like to determine dissociation constant of ovalbumin-anti-OVA-antibody and to compare it with dissociation constant of protein G- immunoglobulin couple. Also, we would like to determine dissociation constants of mumps and measles viruses with their respective antibodies to gain insight into these interactions also.

Izvorni jezik
Engleski

Znanstvena područja
Kemija



POVEZANOST RADA


Projekti:
HRZZ-UIP-2013-11-8193 - Kromatografsko pročišćavanje biomolekula i njihova karakterizacija (CHROBIO) (Brgles, Marija, HRZZ - 2013-11) ( CroRIS)

Ustanove:
Sveučilište u Zagrebu

Profili:

Avatar Url Beata Halassy (autor)

Avatar Url Marija Brgles (autor)

Avatar Url Dubravko Forčić (autor)


Citiraj ovu publikaciju:

Sviben, Dora; Halassy, Beata; Forčić, Dubravko; Brgles, Marija
Impact of dissociation constant on immunoaffinity and affinity chromatography // Symposium & Workshop on Microscale Thermophoresis : book of abstracts / Godinić Mikulčić, Vlatka ; Sviben, Igor ; Rokov Plavec, Jasmina (ur.).
Zagreb: University of Zagreb, Faculty of Science, Chemistry Department, Division of Biochemistry, 2015. str. 31-31 (poster, nije recenziran, sažetak, znanstveni)
Sviben, D., Halassy, B., Forčić, D. & Brgles, M. (2015) Impact of dissociation constant on immunoaffinity and affinity chromatography. U: Godinić Mikulčić, V., Sviben, I. & Rokov Plavec, J. (ur.)Symposium & Workshop on Microscale Thermophoresis : book of abstracts.
@article{article, author = {Sviben, Dora and Halassy, Beata and For\v{c}i\'{c}, Dubravko and Brgles, Marija}, year = {2015}, pages = {31-31}, keywords = {affinity chromatography, immunoaffinity chromatography, dissociation constant}, isbn = {978-953-6076-37-6}, title = {Impact of dissociation constant on immunoaffinity and affinity chromatography}, keyword = {affinity chromatography, immunoaffinity chromatography, dissociation constant}, publisher = {University of Zagreb, Faculty of Science, Chemistry Department, Division of Biochemistry}, publisherplace = {Zagreb, Hrvatska} }
@article{article, author = {Sviben, Dora and Halassy, Beata and For\v{c}i\'{c}, Dubravko and Brgles, Marija}, year = {2015}, pages = {31-31}, keywords = {affinity chromatography, immunoaffinity chromatography, dissociation constant}, isbn = {978-953-6076-37-6}, title = {Impact of dissociation constant on immunoaffinity and affinity chromatography}, keyword = {affinity chromatography, immunoaffinity chromatography, dissociation constant}, publisher = {University of Zagreb, Faculty of Science, Chemistry Department, Division of Biochemistry}, publisherplace = {Zagreb, Hrvatska} }




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