Antibody Fragmentation Service

Specific antigen-binding activity of antibodies has made these molecules the basis of all immunological applications in cell and molecular biology. However, some applications require to study or use only one portion of an immunoglobulin to prevent from unwanted interference with the other portions.

Targeting the hinge region

Distinct fragments of the immunoglobulin can be generated by cleaving the molecule using reducing agents or, more often, using proteases. The hinge region of immunoglobulins is a flexible disulphide-bond rich amino acid portion where the two heavy chains are connected. Targeting the hinge region allows to generate two types of fragments of interest depending on the final application:

  • antigen-binding fragments such as Fab and F(ab’)2
  • class-defining fragments such as Fc which do not bind antigen

Depending on which side of the hinge region (N- or C-terminal) is targeted, the proteolytic cleavage produces either two Fab and one Fc fragment or one F(ab’)2 fragment.

In both cases, an optimisation of the reaction conditions is required and can reveal itself expensive for not as large quantities of antibody fragments as expected. Thanks to our expertise in antibody engineering and protease biology, we are able to provide a robust fragmentation procedure at adapted price for a quantity of antibody as low as 1 milligram.

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Papain - Generating Fab and Fc fragments

Digestion of immunoglobulins by papain results in the separation of on the one hand the Fc region of the heavy chains and on the other hand the two Fab fragments. These fragments can subsequently be used in some very sensitive applications such as ICC.

Learn more about papain digestion.

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Pepsin - Generating F(ab')2 fragments

Digestion of immunoglobulins by pepsin results in the partial degradation of the Fc region of the heavy chains. The Fab fragments are though still connected via the intact hinge region and form a divalent F(ab’)2 fragment which can then be used for capture.

Learn more about pepsin digestion.

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