Isolation of Polyclonal Antibodies to Collagen by Ion Exchange Chromatography on Column with DEAE-Sepharose

Olena Kravchenko, Larisa Oriabinska, Natalia Raksha, Larisa Ivashko, Mariia Abramova

Abstract


Background. Development of new methods for isolation and purification of polyclonal antibodies.

Objective. Analysis of the efficiency of the ion exchange column with DEAE-Sepharose at isolation of polyclonal antibodies to collagen.

Methods. IgG were isolated from precipitated γ-globulin fraction of serum of immunized rabbit by ion exchange chromatography on column with DEAE-Sepharose.

Results. It was determined that the IgG output at volume ratio of 4:3 of starting material and the carrier applied to the column is 1.67 times higher than the ratio of 2:3. It is shown that IgG goes out of the column together with the protein that has a molecular weight of about 57–60 kDa. We found that 5.123 mg of IgG with minor impurities of serum proteins is obtained from 30 cm3 of rabbit’s serum.

Conclusions. The method of ion exchange chromatography on column with DEAE-Sepharose provides a high yield of IgG with few impurities and can be used as a way of pre-treatment of polyclonal antibodies.

Keywords


Polyclonal antibodies; Collagen; Immunization; Ion exchange chromatography; DEAE-Sepharose

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DOI: https://doi.org/10.20535/1810-0546.2017.3.104595

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