Preparation of IgG F(ab’)2 Fraction Using Pepsin Coated Magnetic Particles (Spherotech Cat. # PEPM-40-2)
The Spherotech Cat. No. PEPM-40 consists of 4µm magnetic particles covalently linked to pepsin from porcine stomach mucosa. These particles efficiently cleave IgG from various species producing F(ab’)2 and other lower molecular weight products within a few hours. After digestion Pepsin-Magnetic particles can be easily removed magnetically from the reaction vessel leaving no pepsin in the solution. As a result, the cleavage of IgG can be effectively controlled. The supernatant containing products from the cleavage and any residual IgG can be further purified chromatographically or by using Protein A magnetic particles (Spherotech Cat. # PAMX-10).
Preparation of IgG Fab Fraction Using Papain Coated Magnetic Particles (Spherotech Cat. # PAPM-40-2)
The Spherotech PAPM-40 consists of magnetic particles (4µm) covalently linked to Papain from Papaya latex.These particles efficiently cleave IgG into Fab and Fc fragments within a few hours. After digestion Papain-Magnetic particles can be easily removed from the reaction vessel magnetically leaving no Papain in the solution. As a result, effectively stopping further cleavage of IgG. The supernatant containing products of cleavage and any undigested IgG can be further purified chromatographically.
This procedure allows the controlled preparation of essentially pure Fab in simple two steps. In addition, papain magnetic particles can be reused repeatedly without a significant loss in activity.
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