Thermostabilization of protective antigen--the binding component of anthrax lethal toxin

Journal of Biotechnology
C RadhaRakesh Bhatnagar

Abstract

Protective antigen (PA) is the binding component of anthrax lethal toxin produced by Bacillus anthracis, and constitutes a major ingredient of the vaccine against anthrax. PA and lethal factor when added together are cytolytic to mouse macrophages and J774G8 macrophage cell line. This in vitro lethal toxicity assay is very useful in understanding the molecular mechanism of action of lethal toxin. Effective utilization of PA is, however, hampered due to its thermolability. On prolonged storage at 37 degrees C, PA was found to lose its activity almost completely. The effect of solvent additives like trehalose, sorbitol, xylitol, sodium citrate and magnesium sulphate on the thermal stabilization of PA was examined. The results indicated an increase in the stability of PA when the incubation at 37 degrees C was carried out in the presence of solvent additives used in the 1-3 M range. Magnesium sulphate helped retain the activity up to 82.7% against the control in which no additive was used, as judged by cytolytic assay using J774G8 macrophage cell line. Trehalose or sodium citrate also showed an appreciable protection of PA activity, while sorbitol or xylitol were not very effective. Competitive binding assay using radiolabeled PA ...Continue Reading

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Citations

Mar 17, 2004·Biochemical and Biophysical Research Communications·Samer SinghRakesh Bhatnagar
Nov 25, 2004·Progress in Biophysics and Molecular Biology·Michael W Parker, Susanne C Feil
Sep 5, 2002·Biochemical and Biophysical Research Communications·Samer SinghRakesh Bhatnagar
Dec 14, 1999·Microbes and Infection·S F Little, B E Ivins
Mar 3, 2012·Molecular Pharmaceutics·L K PetersenB Narasimhan
Dec 15, 2010·Clinical and Vaccine Immunology : CVI·Juan Castelán-VegaJuan Arciniega
May 1, 2001·Biochemical and Biophysical Research Communications·V ChauhanR Bhatnagar
Aug 4, 2001·Biochemical and Biophysical Research Communications·N AhujaR Bhatnagar
Jun 9, 2001·Biochemical and Biophysical Research Communications·P GuptaR Bhatnagar
Jul 30, 1999·Protein Expression and Purification·P GuptaR Bhatnagar
Sep 1, 2004·Biochemical and Biophysical Research Communications·Samer SinghRakesh Bhatnagar
Nov 30, 2005·Journal of Pharmaceutical Sciences·Ge JiangVincent J Sullivan
Sep 29, 2005·The Journal of Biological Chemistry·Gil ZomberEytan Elhanany
Jul 31, 2020·Expert Opinion on Biological Therapy·Manish ManishRakesh Bhatnagar
Dec 16, 2003·Biochemical and Biophysical Research Communications·Joungmok KimMoon-Young Yoon

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