Zahra Moradi Nebrin
1, Jafar Majidi
2*, Leili Aghebati Maleki
2, Tohid Kazemi
2, Jalal Abdolalizadeh
3, Somayeh Dadashi
2, Sadeg Eivazi
2, Majid Ahmadi
2, Naeemeh Majidi Zolbanin
41 International University of Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran
2 Departments of Immunology, School of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
3 Immunology Laboratory, Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz-Iran
4 Pharmacology Departments, School of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran
Abstract
Background: Purified mouse IgG2a (a product that could be used in medical research) subclass could be used for animal immunization to production of polyclonal Antibody and to obtain hybridomas in Monoclonal Antibody Production Procedures. The goal of this study was to purify the mouse IgG2a.
Methods: In one step, Ion exchange chromatography was carried out for purification of mouse IgG and then in second step, ProA affinity chromatography was used for IgG2a purification .The chosen method for determination of purity was reduced and non-reduced SDS-PAGE. ELISA method was used for titer and isotype determination.
Results: Mouse Igs with a protein concentration of 27mg/ml (volume: 3CC) was applied on Ion exchange column. Purification by Ion exchange chromatography yielded about 28mg of mouse IgG. Eight mg mouse IgG2a was obtained by ProA affinity chromatography. In reduced SDS-PAGE analysis of purified antibody, two bands were seen in 25& 50 KDa MW positions. Isotype determination of purified mouse IgG2a with mouse isotyping Kit showed the presence of mouse IgG2a isotype with a kappa light chain in related fraction.
Conclusion: Purified mouse IgG2a subclass was obtained with purity more than 95%. Due to the obtained high purity we concluded that Ion exchange chromatography following by ProA affinity chromatography could be a suitable method for purification of mouse IgG subclasses with high quality. Our product is an economical and suitable product that takes a step towards self-sufficiency of the country.