Purification of equine IgG using membrane based enhanced hybrid bioseparation technique: a potential method for manufacturing hyperimmune antibody.

The research paper discusses an efficient method to purify equine immunoglobulin G (IgG) – a hyperimmune antibody used in the production of antivenom and anti-rabies substances – from horse serum using a membrane based enhanced hybrid bioseparation technique. This improved technique significantly increases both purity and recovery rate of equine IgG compared to currently used purification methods.

Technique and Methodology

Within this research, a membrane-based enhanced hybrid bioseparation technique was utilised for the purification of equine IgG, using the following key procedures:

• Firstly, equine IgG is selectively captured from horse serum and retained within a stirred cell membrane module. This is achieved by the addition of an antichaotropic salt which promotes the precipitation and microfiltration of IgG, as well as facilitating hydrophobic interaction based membrane adsorption.
• Any impurities present are subsequently washed out of the membrane module, leaving only the required equine IgG.
• The captured IgG is then released from the stirred cell by reducing the salt concentration. This assists both in the dissolution of the precipitated IgG and in the desorption of the IgG which is bound to the membrane.

Modifications to the Standard Process

The technique employs specific modifications to the standard hybrid bioseparation process:

• Enhancement of IgG recovery from the membrane module is enabled by a switch from a stirring mode to a non-stirring one during the IgG releasing phase.
• Membrane fouling is significantly reduced using a pH switch, thereby improving the overall efficiency of the process.
• To remove ammonium sulfate and lower molecular weight impurities, the enhanced hybrid bioseparation technique is followed by an ultrafiltration step.

Before its practical application, the effect of various operating conditions on equine IgG capture was studied meticulously, ensuring the optimization of the technique.

Results and Significance

The results of applying this updated technique were noteworthy:

• Under optimized conditions, the reaped equine IgG had a purity of 88%.
• The recovery rate of the IgG was over 90%, which was a significant contribution compared to values obtained from traditional purification techniques.

These findings suggest that the membrane-based enhanced hybrid bioseparation technique offers a promising, efficient, and scalable solution for the manufacturing of hyperimmune antibodies.