Rhinogen ^® IdeS protease is a recombinant IdeS protease produced by recombinant expression in the E. coli expression system and obtained through molecular modification. Its molecular weight is about 36kDa. It cleaves IgG only at a specific site below the hinge region to produce F(ab')2 and Fc fragments. It cleaves IgG under physiological conditions to maintain immunogenicity. In common buffers in the pH range of 6.0~8.0, IdeS protease can effectively cleave a variety of antibodies. The modified IdeS protease has higher enzymatic activity and wider substrate specificity. In addition to cleaving human IgG1~4 (cleavage sites are shown in Table 1 below), monkey, sheep, and rabbit IgG, it also has specific cleavage activity for mouse IgG2a and IgG3.

Rhinogen ^® Immobilized IdeS , Microspin is a resin that covalently couples Rhinogen ^® IdeS protease to agarose beads, which cleaves IgG at specific sites to generate F(ab')2 and Fc fragments. After digestion, F(ab')2 and Fc fragments are obtained in the solution without IdeS enzyme.

Packing Specifications:

Rhinogen ^® Immobilized IdeS , Microspin packaging specifications are as follows:

One Immobilized IdeS Microspin can be used to digest 0.5 mg IgG.

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

Storage conditions:

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

Features:

Rhinogen ^® Immobilized IdeS , Microspin has the following characteristics:

1. Easy to use: After digestion, the digestion products can be separated by centrifugation;

2. High recovery rate: F(ab')2 recovery rate>85%;

3. 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.

Instructions:

1. Rhinogen ^® Immobolized IdeS , Microspin can be effectively used for enzymatic cleavage of human, humanized, chimeric, sheep, rabbit and monkey IgG, and also has moderate activity against mouse IgG2a and IgG3. It is also suitable for the enzymatic cleavage of many Fc-fusion proteins and antibody drug conjugates (ADCs); it is not suitable for mouse IgG1/IgG2b, rat, pig, cow or goat IgG. In addition, it can cleave 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 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. IgG cleavage can be easily determined by SDS-PAGE;

5. After incubating the digestion product with protein A magnetic beads for 30 to 60 minutes, the purified F(ab ' )2 fragment can be obtained. RhinoBio also provides the product Rhinogen ^® Immobolized IdeS Cut-Pure Kit, Microspin (Cat. No.: QIP-102), which contains Immobilized IdeS , Microspin and Protein A purification column;

6. The optimal pH of the enzyme digestion buffer is 6.0-8.0. In a system below pH 5.0, the IdeS immobilized enzyme will be irreversibly inactivated. In a system above pH 8.0, the reaction conditions need to be appropriately extended/optimized.

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