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An easy and simple separation method for Fc and Fab fragments from chicken immunoglobulin Y (IgY)
- Source :
- Journal of Chromatography B. 1141:122011
- Publication Year :
- 2020
- Publisher :
- Elsevier BV, 2020.
-
Abstract
- Antigen-binding (Fab) and crystallizable (Fc) fragments are the active components of yolk immunoglobulin (IgY), which have been widely used in the pharmaceutical field. However, the common purification methods for the Fab and Fc fragments use combinations of multi-columns are complex and time-consuming. The objective of this study was to improve the separation efficiency of the Fab and Fc fragments from the hydrolyzed IgY and increase the purity of the isolated Fab and Fc fragments. Natural IgY was hydrolyzed using papain for 6 hr and then treated with 45% saturated ammonium sulfate to remove small molecular-weight-peptides. The fraction containing Fab and Fc fragments was loaded on a DEAE-Sepharose ion exchange column and the Fab fraction was washed out first with 10 mM Tris-HCl buffer (pH 7.6). Then, the Fc fraction bound to the DEAE Sepharose was eluted with 10 mM Tris-HCl buffer (pH 7.6) containing 0.21 M NaCl. The purity of the two fragments was 88.7% and 90.1%, respectively. The results of Western blotting and MS analyses indicated that this method purified Fab and Fc fractions with high purity. This method is easy and simple compared with other methods, and the active fragments separated can be easily used.
- Subjects :
- Ammonium sulfate
Blotting, Western
Clinical Biochemistry
Immunoglobulins
Fraction (chemistry)
030226 pharmacology & pharmacy
01 natural sciences
Biochemistry
Analytical Chemistry
Immunoglobulin Fab Fragments
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
Papain
Animals
Chromatography
Ion exchange
biology
Chemistry
Elution
010401 analytical chemistry
Cell Biology
General Medicine
Chromatography, Ion Exchange
Immunoglobulin Fc Fragments
0104 chemical sciences
DEAE-Sepharose
Ammonium Sulfate
biology.protein
Immunoglobulin Y
Antibody
Chickens
Subjects
Details
- ISSN :
- 15700232
- Volume :
- 1141
- Database :
- OpenAIRE
- Journal :
- Journal of Chromatography B
- Accession number :
- edsair.doi.dedup.....c89226b35cadebc722f74560e7989138