background:

IdeS protease from Streptococcus pyogenes is an immunoglobulin degrading enzyme that cleaves IgG only at a specific site below the hinge region to produce F(ab')2 and Fc fragments. Since IdeS protease digests IgG at a specific site, there is no risk of over-digestion by extending the incubation time.

Rhinogen ^® Immobolized IdeS Cut-Pure Kit, Microspin contains Immobilized IdeS , Microspin for antibody digestion and Protein A purification column for separation of F(ab')2 fragments and Fc fragments.

Rhinogen ^® Immobilized IdeS , Microspin uses a modified IdeS protease, so it has a wider substrate specificity and can effectively cleave human IgG1~4 (cleavage sites are shown in Table 1 below), monkey, sheep, and rabbit IgG, and can also specifically cleave mouse IgG2a and IgG3. In addition, Immobilized IdeS , Microspin can cleave IgG under physiological conditions, thereby maintaining the immunogenicity of the enzymatic cleavage product.

The Protein A purification column uses recombinantly expressed Protein A, which has enhanced the specificity of binding to Fc and improved protein stability compared to natural Protein A. The Protein A purification column can specifically bind to the Fc segment of a variety of immunoglobulins.

Packing Specifications:

Rhinogen ^® Immobolized IdeS Cut-Pure Kit, Microspin packaging specifications are as follows:

A set of Immobilized IdeS , Microspin and Protein A purification columns can be combined for enzymatic digestion and purification of 0.5 mg IgG .

The product is stored in 20% ethanol without added preservatives.

Storage conditions:

Transported with ice packs, Immobilized IdeS , Microspin and Protein A purification column products are stored in 20% ethanol. Please store them at 2-8℃ immediately after receiving the products. Avoid freezing!

Features:

Rhinogen ^® Immobolized IdeS Cut-Pure Kit, Microspin has the following features:

1. High specific activity : effectively cut the lower hinge of the antibody to obtain F(ab ' )2 and Fc fragments; the enzyme cleavage efficiency is ≥90%;

2. Broad substrate specificity: can effectively cut human IgG1~4, monkey, sheep, rabbit IgG, mouse IgG2a, IgG3;

3. High recovery rate: After digestion, purification and separation, F(ab')2 can be harvested with a recovery rate of more than 80%;

4. High stability: Each batch of products undergoes strict quality control to achieve product batch-to-batch stability .

application:

Structural characterization and analysis of recombinant antibody drugs, preparation of Fab fragments.

Instructions:

1. Rhinogen ^® Immobolized IdeS Cut-Pure Kit, Microspin can be effectively used for enzymatic cleavage and purification of human, humanized, chimeric, sheep, rabbit and monkey IgG, and has moderate activity against mouse IgG2a and IgG3. It is also suitable for the enzymatic cleavage and purification of many Fc-fusion proteins and antibody drug conjugates (ADCs) . It is not suitable for mouse IgG1/IgG2b, rat, pig, cattle or goat IgG. In addition, it cannot cut non- IgG isotypes, including IgA, IgM, IgD and IgE ;

2. For the cleavage of mouse IgG2a and mouse IgG3, it is recommended to moderately increase the incubation temperature (the closer the enzyme cleavage temperature is to 37°C, the higher the enzyme activity ) and incubation time (2 hours to overnight);

3. The ideal IgG concentration should be in the range of 0.5~20mg/ml;

4. The cleavage of the target protein can be easily determined by SDS-PAGE;

5. The optimal pH of the enzyme digestion buffer is 6.0~8.0. In a system below pH 5.0, Immobolized IdeS will be irreversibly inactivated, and the reaction conditions need to be appropriately prolonged/optimized in systems above pH 8.0;

6. This product is for research use only and is not intended for diagnosis or treatment of humans or animals.