Optimization of Unnicked β2-Glycoprotein I and High Avidity Anti- β2-Glycoprotein I Antibodies Isolation
Joint Authors
Ambrožič, Aleš
Artenjak, Andrej
Leonardi, Adrijana
Križaj, Igor
Božič, Borut
Čučnik, Saša
Sodin-Semrl, Snezna
Source
Journal of Immunology Research
Issue
Vol. 2014, Issue 2014 (31 Dec. 2014), pp.1-8, 8 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2014-01-23
Country of Publication
Egypt
No. of Pages
8
Main Subjects
Abstract EN
Patient biological material for isolation of β2-glycoprotein I (β2GPI) and high avidity IgG anti-β2-glycoprotein I antibodies (HAv anti-β2GPI) dictates its full utilization.
The aim of our study was to evaluate/improve procedures for isolation of unnicked β2GPI and HAv aβ2GPI to gain unmodified proteins in higher yields/purity.
Isolation of β2GPI from plasma was a stepwise procedure combining nonspecific and specific methods.
For isolation of polyclonal HAv aβ2GPI affinity chromatographies with immobilized protein G and human β2GPI were used.
The unknown protein found during isolation was identified by liquid chromatography electrospray ionization mass spectrometry and the nonredundant National Center for Biotechnology Information database.
The average mass of the isolated unnicked purified β2GPI increased from 6.56 mg to 9.94 mg.
In the optimized isolation procedure the high molecular weight protein (proteoglycan 4) was successfully separated from β2GPI in the 1st peaks with size exclusion chromatography.
The average efficiency of the isolation procedure for polyclonal HAv anti-β2GPI from different matrixes was 13.8%, as determined by our in-house anti-β2GPI ELISA.
We modified the in-house isolation and purification procedures of unnicked β2GPI and HAv anti-β2GPI, improving the purity of antigen and antibodies as well as increasing the number of tests routinely performed with the in-house ELISA by ~50%.
American Psychological Association (APA)
Artenjak, Andrej& Leonardi, Adrijana& Križaj, Igor& Ambrožič, Aleš& Sodin-Semrl, Snezna& Božič, Borut…[et al.]. 2014. Optimization of Unnicked β2-Glycoprotein I and High Avidity Anti- β2-Glycoprotein I Antibodies Isolation. Journal of Immunology Research،Vol. 2014, no. 2014, pp.1-8.
https://search.emarefa.net/detail/BIM-1040787
Modern Language Association (MLA)
Artenjak, Andrej…[et al.]. Optimization of Unnicked β2-Glycoprotein I and High Avidity Anti- β2-Glycoprotein I Antibodies Isolation. Journal of Immunology Research No. 2014 (2014), pp.1-8.
https://search.emarefa.net/detail/BIM-1040787
American Medical Association (AMA)
Artenjak, Andrej& Leonardi, Adrijana& Križaj, Igor& Ambrožič, Aleš& Sodin-Semrl, Snezna& Božič, Borut…[et al.]. Optimization of Unnicked β2-Glycoprotein I and High Avidity Anti- β2-Glycoprotein I Antibodies Isolation. Journal of Immunology Research. 2014. Vol. 2014, no. 2014, pp.1-8.
https://search.emarefa.net/detail/BIM-1040787
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1040787