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Immunoaffinity purification of albumin from fish serum

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Date Issued:
1998
Summary:
This study describes an affinity method which purifies albumin from fish serum. Hybridoma clones producing antibodies specific to fish albumin have been screened and propagated and their antibodies coupled to a cross-linked agarose gel matrix which has been activated with N-hydroxysuccinimide. The resulting gel with its ligand attached was then used to purify albumin from fish serum. The procedure was optimized for coupling, sample application and elution. The coupling optimization using varying pH values resulted in over 90% coupling efficiencies. The sample application optimization showed yields of 2 mg of protein per ml of serum at 1:5 dilution of sample. Elution conditions were evaluated using varying pH and chaotropic agent concentrations. The optimal elution conditions (pH 4.0) resulted in a yield of 0.2 mg/ml of gel with several impurities. Elution at pH 3.0 resulted in a very pure product with low yield (.02 mg/ml of gel) and a loss of activity.
Title: Immunoaffinity purification of albumin from fish serum.
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Name(s): Apter, Christine V.
Florida Atlantic University, Degree grantor
Hartmann, James X., Thesis advisor
Type of Resource: text
Genre: Electronic Thesis Or Dissertation
Issuance: monographic
Date Issued: 1998
Publisher: Florida Atlantic University
Place of Publication: Boca Raton, Fla.
Physical Form: application/pdf
Extent: 51 p.
Language(s): English
Summary: This study describes an affinity method which purifies albumin from fish serum. Hybridoma clones producing antibodies specific to fish albumin have been screened and propagated and their antibodies coupled to a cross-linked agarose gel matrix which has been activated with N-hydroxysuccinimide. The resulting gel with its ligand attached was then used to purify albumin from fish serum. The procedure was optimized for coupling, sample application and elution. The coupling optimization using varying pH values resulted in over 90% coupling efficiencies. The sample application optimization showed yields of 2 mg of protein per ml of serum at 1:5 dilution of sample. Elution conditions were evaluated using varying pH and chaotropic agent concentrations. The optimal elution conditions (pH 4.0) resulted in a yield of 0.2 mg/ml of gel with several impurities. Elution at pH 3.0 resulted in a very pure product with low yield (.02 mg/ml of gel) and a loss of activity.
Identifier: 9780591929850 (isbn), 15568 (digitool), FADT15568 (IID), fau:12328 (fedora)
Collection: FAU Electronic Theses and Dissertations Collection
Note(s): Charles E. Schmidt College of Science
Thesis (M.S.)--Florida Atlantic University, 1998.
Subject(s): Serum albumin--Purification
Immune serums
Held by: Florida Atlantic University Libraries
Persistent Link to This Record: http://purl.flvc.org/fcla/dt/15568
Sublocation: Digital Library
Use and Reproduction: Copyright © is held by the author, with permission granted to Florida Atlantic University to digitize, archive and distribute this item for non-profit research and educational purposes. Any reuse of this item in excess of fair use or other copyright exemptions requires permission of the copyright holder.
Use and Reproduction: http://rightsstatements.org/vocab/InC/1.0/
Host Institution: FAU
Is Part of Series: Florida Atlantic University Digital Library Collections.