Igg Purification

II. INTRODUCTION

Blood plasma contain blood serum which is component of the plasma that does not contain any cells or clotting factors. Protein found there are called serum proteins. These proteins carry out a number of functions in organisms such as they act as circulatory transport molecules, enzymes, and participate in the proper functioning of the immune system. Serum proteins are largely consisted of albulmin, immunoglobulin and fibrinogen. Immunoglobulins are composed of α, β, and γ types. Each consists of two large heavy chains (50 kDa to 55 kDa) and two light chains with molecular weight (22.5 kDa to 25 kDa) (West and Hong, 1962).Certain diseases, such as acute infection, hyperimmunization and multiple myeloma, may be associated with an increase in gamma globulin proteins (reference). Globulin proteins might also be linked to Hepatitis C virus (refer). Serum globulin electrophoresis is often conducted as a medical laboratory test to assess the concentration of gamma globulin components (M, G and A). Thus IgG purification and quantitation can allow research and development in diagnosing diseases.

Immunoglobulin G (IgG) has pI ranges from 5.8 to 7.3. Ammonium sulfate is used to precipitate the IgG proteins from the serum via disrupting the protein interaction with the water molecules (zhao). Thus the proteins will aggregate and precipitated. Due to the pI value of IgG, it can be excluded from other serum proteins via passing through an anion exchanger. Finally Sodium Dodecyl Sulfate – Polyacrylamid gel can be used to separate the proteins according to their molecular weight differences. (Reference)

An experiment was carried out to fractionate the rabbit serum into different fractions and purify IgG globulin proteins, and evaluate the efficiency of this procedure which involved ammonium sulfate precipitation, DEAE weak anion exchanger and 12% SDS-PAGE methods.

III. MATERIALS AND METHODS

III. a) Precipitation Of Serum Globulins Using Ammonium Sulphate...