eISSN: 1644-4124
ISSN: 1426-3912
Central European Journal of Immunology
Current issue Archive Manuscripts accepted About the journal Special Issues Editorial board Abstracting and indexing Subscription Contact Instructions for authors Publication charge Ethical standards and procedures
Editorial System
Submit your Manuscript
SCImago Journal & Country Rank
Search
1/2014
vol. 39
 
Share:
Send email
Share:
abstract:

Purification of pituitary autoantigen by column liquid chromatography and chromatofocusing

Paweł Gut
,
Jakub Fischbach
,
Katarzyna Ziemnicka
,
Maciej Bączyk
,
Daria Baszko-Błaszyk
,
Elżbieta Wrotkowska
,
Marek Ruchała

(Centr Eur J Immunol 2014; 39 (1): 46-50)
DOI: https://doi.org/10.5114/ceji.2014.42122
Online publish date: 2014/04/17
View full text Get citation
 
PlumX metrics:
Introduction: Pituitary autoantibodies can be determined both in patients with pituitary disease as well as patients with autoimmune endocrine diseases. The purpose of the study was to isolate and purify pituitary autoantigen using sera of patients and the microsomal fraction of the pituitary.

Material and methods: To isolate a pituitary autoantigen, patient sera were used, which showed a strong immune response to pituitary antigens. Pituitary microsomal fractions were prepared from pituitary tissue homogenates. In the study, sera of patients with pituitary disease, Addison and Graves’ disease were used. The initial stages were carried out by affinity chromatography on CN-Br sepharose column whereas purification was continued by column liquid chromatography on AcA54 Ultrogel. Chromatofocusing was performed by Polybuffer exchanger PBE 94.

Results: 125I-labeled pituitary antigens after isolation appeared in column chromatography in three peaks. The first peak contained 50-70 kDa proteins, the second peak – 17 to 22 kDa proteins and the third peak contains 125-iodides. Three fractions obtained from filtration on Ultrogel were separated in a polyacrylamide gel. In the first peak two bands 67 and 55 kDa appeared. The second peak contained low molecular weight substances, and the third peak contained 125I. The first peak from Ultrogel was isolated by chromatofocusing – the first peak with pH 5.9 and the second one with pH 4.9.

Conclusions: Isolation and purification of pituitary autoantigen with the use of column liquid chromatography and chromatofocusing resulted in obtainment of two antigenic proteins of specific gravity of 67 and 55 kDa.
keywords:

pituitary autoantigen, purification, column liquid chromatography, chromatofocusing
Termedia
About us Contact
Copyright notice Privacy policy Advertising policy Contact us
Quick links
Journals Books eBooks Events
© 2024 Termedia Sp. z o.o.
Developed by Bentus.