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International Journal of Medical Laboratory
Shahid Sadoughi University of Medical Sciences
Wed, Apr 24, 2024
[Archive]
Volume 7, Issue 1 (February 2020)
IJML 2020, 7(1): 49-58 |
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Kordzangene A, Mohebat R, Mosslemin M H, Taghavi Moghadam A. Reduction of Purification Time of Polyspecific Equine F(ab´)2 Antivenom against Scorpion Envenomation. IJML 2020; 7 (1) :49-58
URL: http://ijml.ssu.ac.ir/article-1-314-en.html
URL: http://ijml.ssu.ac.ir/article-1-314-en.html
Department of Chemistry, Yazd Branch, Islamic Azad University, Yazd, Iran.
Abstract: (1031 Views)
Background and Aims: In this study we improved the purification of immunoglobulins from equine antiserum raised against the venom of 6 types of scorption species. Caprylic acid (octanoic acid), a fatty acid, was found to have no effect on the activity of the enzymes pepsin, which is used in antivenom purification to digest Fc fragment of immunoglobulins to obtain F(ab´)2.
Materials and Methods: A new method was developed for the production of F(ab´)2 antivenom whereby whole equine antiserum was mixed with equal amount of 0.15 M HCl and pH 3.4 with pepsin 660 mg/L of diluted antivenom and incubated for 4 h at 37°C. After digestion the pH were brought to 4.8 with sodium hydroxide solution (1.5 M) and then 1.5% caprylic acid and 10% ammonium sulfate was added and mixed for 60 minutes and passed through filter paper.
Results: Caprylic acid caused precipitation of albumin, and ammonium sulfate reduced turbidity of solution, resulting in a reduced protein load presented to the digestion enzymes culminating in substantial reductions in processing time.
Conclusions: The equine F(ab´)2 obtained using these novel caprylic acid methods were comparable in terms of yield, purity and specific activity to those obtained by multi-step and time consuming conventional salt fractionation with ammonium sulfate.
Materials and Methods: A new method was developed for the production of F(ab´)2 antivenom whereby whole equine antiserum was mixed with equal amount of 0.15 M HCl and pH 3.4 with pepsin 660 mg/L of diluted antivenom and incubated for 4 h at 37°C. After digestion the pH were brought to 4.8 with sodium hydroxide solution (1.5 M) and then 1.5% caprylic acid and 10% ammonium sulfate was added and mixed for 60 minutes and passed through filter paper.
Results: Caprylic acid caused precipitation of albumin, and ammonium sulfate reduced turbidity of solution, resulting in a reduced protein load presented to the digestion enzymes culminating in substantial reductions in processing time.
Conclusions: The equine F(ab´)2 obtained using these novel caprylic acid methods were comparable in terms of yield, purity and specific activity to those obtained by multi-step and time consuming conventional salt fractionation with ammonium sulfate.
Type of Study: Research |
Subject:
Biochemistry
Received: 2019/05/11 | Accepted: 2019/11/12 | Published: 2020/01/30
Received: 2019/05/11 | Accepted: 2019/11/12 | Published: 2020/01/30
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