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Thermal Unfolding Curves of High Concentration Bovine IgG Measured by FTIR Spectroscopy

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Abstract

The purpose of this research is to study the thermal unfolding of high concentration bovine Immunoglobulin G (IgG) under 26 different experimental conditions by Fourier Transform Infrared spectroscopy with improved purge conditions and software calculations. When bovine IgG (25–200 mg/mL) was thermally denatured between pH 4.0 and 8.0, it was observed that at 25 mg/mL concentration, the protein exhibited maximum thermal stability at pH 6.0 and 7.0 as evident from the apparent Tm values. Increasing the concentration from 25 to 100 mg/mL at those pH values increased the thermal resistance of the protein by 2–3 °C. But, at 200 mg/mL, IgG showed a small decrease in its transition temperature. Presence of 100 mM Trehalose enhanced the Tm values at all conditions and possibly prevented the complete loss of IgG as insoluble aggregates at higher temperatures. Second derivative plots were constructed to explain the conformational changes of IgG during thermal unfolding.

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Abbreviations

FTIR spectroscopy:

Fourier transform infrared spectroscopy

Immunoglobulin G:

IgG

CD:

Circular dichroism

DSC:

Differential scanning calorimetry

NMR:

Nuclear magnetic resonance

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Author notes

  1. V. Sathya Devi

    Present address: Integrated BioTherapeutics Inc., 21 Firstfield Road, Gaithersburg, MD, 20878, USA

  2. Jeremy Truntzer

    Present address: UC Irvine School of Medicine, Irvine, CA, 92617, USA

Authors and Affiliations

  1. Coleman Softlabs, Inc., 296 Bay Road, Atherton, CA, 94027, USA

    V. Sathya Devi, Denis R. Coleman & Jeremy Truntzer

Authors
  1. V. Sathya Devi
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  2. Denis R. Coleman
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  3. Jeremy Truntzer
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Correspondence to V. Sathya Devi or Denis R. Coleman.

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Sathya Devi, V., Coleman, D.R. & Truntzer, J. Thermal Unfolding Curves of High Concentration Bovine IgG Measured by FTIR Spectroscopy. Protein J 30, 395–403 (2011). https://doi.org/10.1007/s10930-011-9344-y

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