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Studies on the fractionation of Equine Antivenom IgG by combinations of Ammonium sulfate and Caprylic acid.
MedLine Citation:
PMID:  22842065     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
This study involved the use of combined stepwise ammonium sulfate (AS) and caprylic acid (CA) fractionation of equine antivenom IgG without intermediate separation of precipitate. Using a microplate and checker board titration format, plasma was treated under 66 conditions with varying concentrations of AS (0-25% saturation) and CA (0-5% v/v). The filtrate of each well was assayed for protein and antibody activity. At about 1.5 - 4.0% CA without AS, the precipitated plasma gave high specific antibody activity. Twelve precipitation conditions selected from the microplate experiment were studied in detail in tubes. The highest turbidity was with 5% CA alone. The highest antibody recovery of 95.45% was observed at 15% AS with 3.0% CA. The highest specific activity with 3.28 folds purification was observed with 4.0% CA. Thus, AS could reduce the turbidity induced by CA and increase the yield but not the purity of antibody. Size exclusion HPLC showed the antibody to be one single peak with 1.5% of soluble protein aggregate. When all parameters were considered, the optimum fractionation condition appeared to be 3.5% CA alone which gave high specific antibody activity (3.26 folds purification), antibody recovery (93.93%) and low turbidity (0.56% solid). Furthermore, better overall results were observed with one hour than overnight precipitation.
Authors:
Sukanya Eursakun; Pavinee Simsiriwong; Kavi Ratanabanangkoon
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-7-26
Journal Detail:
Title:  Toxicon : official journal of the International Society on Toxinology     Volume:  -     ISSN:  1879-3150     ISO Abbreviation:  -     Publication Date:  2012 Jul 
Date Detail:
Created Date:  2012-7-30     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  1307333     Medline TA:  Toxicon     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2012. Published by Elsevier Ltd.
Affiliation:
Chulabhorn Graduate Institute; Center of Excellence on Environmental Health and Toxicology (EHT).
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