Your browser does not allow JavaScript!
JavaScript is necessary for the proper functioning of this website. Please enable JavaScript or use a modern browser.
Open Science Slovenia
Open Science
DiKUL
slv
|
eng
Search
Browse
New in RUL
About RUL
In numbers
Help
Sign in
Noninvasive method for determination of immobilized protein A
ID
Mravljak, Rok
(
Author
),
ID
Stantič, Metka
(
Author
),
ID
Bizjak, Ožbej
(
Author
),
ID
Podgornik, Aleš
(
Author
)
PDF - Presentation file,
Download
(1,13 MB)
MD5: 0372A609895695A9A73FCEB90A1FD0F1
URL - Source URL, Visit
https://www.sciencedirect.com/science/article/pii/S0021967322001741
Image galllery
Abstract
The pH transition method, developed for the determination of the ion-exchange group density on chromatographic stationary phase, was used for the quantification of immobilized protein A. Monolithic epoxy polyHIPE and particulate CNBr-Sepharose supports were used for immobilization. A lactate buffer was selected, having a buffer capacity peak approximately 0.5 pH units below the maximum buffer capacity of protein A. The pH transition measurements were performed at pH 4.3, where protein A exhibits maximum buffer capacity, with a lactate buffer concentration of 1 mM for protein A immobilized on polyHIPE monoliths and of 5 mM for protein A immobilized on CNBr-Sepharose. The pH transition height and full width at half maximum for the particulate support and the height for the polyHIPE matrix, showed a linear correlation with the amount of immobilized protein A determined from the absorbance difference before and after immobilization for both supports. The developed method allows a simple, non-invasive on-line determination of immobilized protein A using biological buffers, even for chromatographic columns with an amount of immobilized protein A as low as 0.25 mg. In addition, its sensitivity and duration can be easily adjusted by varying the buffer concentration and pH.
Language:
English
Keywords:
direct noninvasive immobilized protein
,
quantification
,
protein A
,
pH transition method
,
polyHIPE
,
CNBr-Sepharose
Work type:
Article
Typology:
1.01 - Original Scientific Article
Organization:
FKKT - Faculty of Chemistry and Chemical Technology
Publication status:
Published
Publication version:
Version of Record
Year:
2022
Number of pages:
7 str.
Numbering:
Vol. 1671, art. 462976
PID:
20.500.12556/RUL-137650
UDC:
66.08:577.112
ISSN on article:
0021-9673
DOI:
10.1016/j.chroma.2022.462976
COBISS.SI-ID:
103441667
Publication date in RUL:
24.06.2022
Views:
607
Downloads:
112
Metadata:
Cite this work
Plain text
BibTeX
EndNote XML
EndNote/Refer
RIS
ABNT
ACM Ref
AMA
APA
Chicago 17th Author-Date
Harvard
IEEE
ISO 690
MLA
Vancouver
:
Copy citation
Share:
Record is a part of a journal
Title:
Journal of chromatography. A
Shortened title:
J. chromatogr., A
Publisher:
Elsevier
ISSN:
0021-9673
COBISS.SI-ID:
6398471
Licences
License:
CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
Link:
http://creativecommons.org/licenses/by-nc-nd/4.0/
Description:
The most restrictive Creative Commons license. This only allows people to download and share the work for no commercial gain and for no other purposes.
Secondary language
Language:
Slovenian
Keywords:
direktna neinvazivna imobilizacija proteina
,
kvantifikacija
,
protein A
,
metoda pH prehoda
,
poliHIPE
,
CNBr-sefaroza
Projects
Funder:
ARRS - Slovenian Research Agency
Project number:
J2-9440
Name:
Karakterizacija fraktalnih struktur in povečevalni kriteriji njihove sinteze
Funder:
ARRS - Slovenian Research Agency
Project number:
P1-0153
Name:
Raziskave in razvoj analiznih metod in postopkov
Similar documents
Similar works from RUL:
Similar works from other Slovenian collections:
Back