FabRICATOR
FabRICATOR is a unique proteolytic enzyme that cleaves the IgG just below the hinge region thereby generating an intact F(ab')2 fragment and a Fc fragment. The enzyme is a modified cysteine protease first isolated from Streptococcus pyogenes. The scientific name of the enzyme is IdeS (immunoglobulin degrading enzyme) which has a biological role of circumventing the host defense.
Advantages of using FabRICATOR for F(ab')2 or Fab' production
- Fast - IgG cleaved within 30 minutes
- No optimization - works right out of the box
- Homogenous - cleavage position exactly the same in each generated fragment
- Gentle method - reaction carried out at physiological pH with no negative effect on immunoreactivity of the generated fragments
Antibodies compatible with FabRICATOR enzyme
- Human IgG
- Humanized IgG
- Chimeric IgG
- Monkey
- Rabbit
- Sheep
- Mouse IgG2a (for reaction conditions go here )
- Mouse IgG3
F(ab')2 Fragments in Minutes
- Molecular weight and marker Mark 12 (Invitrogen)
- IgG fraction
- 5 min incubation with FabRICATOR®
- 15 min incubation with FabRICATOR®
- 30 min incubation with FabRICATOR®
- 30 min incubation with FabRICATOR® (reduced sample)
- After removal of Fc fragments
- After removal of FabRICATOR®
- Molecular weight marker Mark 12
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Fab and F(ab')2 antibody fragments are used in many applications or assays where the presence of the Fc region may cause problems. If antibodies are used for staining of a specific target in tissues like spleen, lymph or in peripheral blood preparations, cells with Fc receptors (macrophages, monocytes, natural killer cells and B lymphocytes) can bind to the Fc region of the antibody and cause a background staining in areas that do not contain any antigen.
Fab and F(ab')2 fragments are also desirable for staining or binding to cell preparations in the presence of plasma since they can not bind to complement, which would lyse the cells. The divalency of the F(ab')2 fragment enables it to cross-link antigens and hence makes it useful for rosetting, cellular aggregation and precipitation assays.
