Article: Novel hydrophobic interaction chromatography matrix for specific isolation and simple elution of immunoglobulins (A, G, and M) from porcine serum.
2006 Volume 1122, Issue 1-2, Page(s) 28–34
Abstract: A new, highly acetylated agarose matrix (HA-Sepharose) was synthesized and used as a hydrophobic interaction chromatography (HIC) medium to specifically isolate immunoglobulins (Igs) from porcine serum. Recovery of Igs was in a single step and under mild ...
| Abstract | A new, highly acetylated agarose matrix (HA-Sepharose) was synthesized and used as a hydrophobic interaction chromatography (HIC) medium to specifically isolate immunoglobulins (Igs) from porcine serum. Recovery of Igs was in a single step and under mild conditions. HA-Sepharose adsorption was studied in terms of salt, gel acetylation time, flow rate, and protein concentration on the loading buffer. At 0.5 M Na2SO4, control with unmodified Sepharose retained a small fraction (0.70 mg/mL of matrix) of serum albumin. On the contrary HA-Sepharose retained primary Igs (IgA, IgG, and 53% of IgM) as revealed by sodium dodecyl sulphate 10% polyacrylamide gel electrophoresis (SDS-PAGE), quantitative radial immunodiffusion and immunodetection. At a flow rate of 1 mL/min, the HA-Sepharose column capacity (3.9 mg/mL of matrix) was similar to the reported capacity for the commercial thiophilic T-gel. However, HA-Sepharose showed higher recovery of IgA and IgM than the T-gel in the same salt conditions, clearly an advantage in terms of immunoglobulin recovery strategies. Acetylation changed the matrix adsorption from albumin to immunoglobulins; thus, the highly acetylated gel rendered recoveries of Igs from unprocessed porcine serum practically free of albumin. |
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| MeSH term(s) | Adsorption ; Animals ; Blotting, Western ; Chromatography, Affinity/methods ; Chromatography, Gel/methods ; Electrophoresis, Polyacrylamide Gel/methods ; Hydrophobic and Hydrophilic Interactions ; Immunoblotting ; Immunoglobulin A/blood ; Immunoglobulin A/isolation & purification ; Immunoglobulin G/blood ; Immunoglobulin G/isolation & purification ; Immunoglobulin M/blood ; Immunoglobulin M/isolation & purification ; Immunoglobulins/blood ; Immunoglobulins/isolation & purification ; Reproducibility of Results ; Sepharose/chemistry ; Swine | |||||
| Chemical Substances | Immunoglobulin A ; Immunoglobulin G ; Immunoglobulin M ; Immunoglobulins ; Sepharose (9012-36-6) | |||||
| Language | English | |||||
| Publishing date | 2006-07-28 | |||||
| Publishing country | Netherlands | |||||
| Document type | Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't | |||||
| ZDB-ID | 1171488-8 | |||||
| ISSN | 1873-3778 ; 0021-9673 | |||||
| ISSN (online) | 1873-3778 | |||||
| ISSN | 0021-9673 | |||||
| DOI | 10.1016/j.chroma.2006.04.012 | |||||
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| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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