background:

IdeS 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. Rhinogen ^® IdeS protease is a recombinant IdeS protein produced by recombinant expression in the E. coli system and obtained through molecular modification . It contains a His tag, which can be easily separated and removed from the enzyme cleavage reaction system after the enzyme cleavage is completed. IdeS protease cleaves IgG under physiological conditions, thereby maintaining immunoreactivity. 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 broader substrate specificity. In addition to cleaving human IgG1~4, monkey, sheep, and rabbit IgG, it also has specific cleavage activity for mouse IgG2a and IgG3.

Packing Specifications:

Rhinogen ^® IdeS protease packaging specifications are as follows:

QIP-001 is a lyophilized powder formulation product with a formulation buffer of 20mM Tris, 50mM NaCl, 1mM EDTA, pH 7.5.

Product Source:

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 approximately 36kDa.

Product Quality:

SDS-PAGE analysis showed purity ≥95%.

Product Features:

The optimum pH is 6.0~8.0.

Enzyme activity definition :

1 unit of enzyme activity is defined as the amount of enzyme required to cleave 1 μg of recombinant monoclonal IgG within 30 minutes at 37°C.

Storage conditions:

1. Use ice packs or dry ice for transportation. Please freeze the product at -30℃ to -10℃ immediately after receiving it;

2. Before use, dissolve the IdeS protease freeze-dried powder with sterile water. After dissolution, store at 2~8℃. If used aseptically , the validity period is 30 days.

Features:

Rhinogen ^® IdeS protease is a highly purified and very stable recombinant protease with high stability and high specific activity. It is suitable for structural characterization analysis of recombinant antibody drugs.

1. High specific activity: effectively cut the lower hinge of the antibody to obtain F(ab ' )2 and Fc fragments;

2. Broad substrate specificity: In addition to the enzymatic activity against human IgG1~4, monkey, sheep, and rabbit IgG , it also has specific cleavage activity against mouse IgG2a and IgG3;

3. No animal origin: Recombinant production, no exogenous virus contamination, and no animal origin raw materials are used in the production process;

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

application:

IgG is cleaved at specific sites to generate F(ab ' )2 and Fc fragments.

Recommended use:

1. Reagent preparation:

1) Before use, please take out the Rhinogen ^® IdeS protease and centrifuge at 10000rpm for 10 seconds to ensure that all the dry powder is at the bottom of the tube;

Rhinogen ^® IdeS protease powder in deionized water to make a 50 units/μl solution.

2. Recommended usage:

1) Add the required amount of IgG (0.5-10 mg/ml) to digestion buffer or other compatible buffer;

2) Add Rhinogen ^® IdeS protease to the IgG sample;

Note: Add 1 unit of IdeS protease for every 1 μg of IgG for digestion; for example, add 1 μl (50 units) of IdeS solution to digest 50 μg of IgG;

3) Incubate at 37°C for 30-60 minutes.

Instructions:

Rhinogen ^® IdeS protease can effectively cleave human, humanized, chimeric, sheep, rabbit and monkey IgG , and has high activity against mouse IgG2a and IgG3. Rhinogen ^® IdeS protease can also cleave many Fc-fusion proteins and antibody drug conjugates (ADCs);

1. IdeS protease cannot cleave mouse IgG1/IgG2b, rat, porcine, bovine or goat IgG. In addition, it cannot cleave non- IgG isotypes, including IgA, IgM, IgD and IgE ;

2. The ideal IgG concentration should be in the range of 0.5~10mg/ml;

3. The pH of the digestion reaction buffer is 6.0-8.0, with an optimum pH of 6.5, and the NaCl concentration should not be too high. The recommended digestion buffer is 10mM sodium phosphate, 10mM NaCl, pH 6.5;

4. For cleavage of mouse IgG2a and mouse IgG3, increase the amount of IdeS protease (5-fold to 10-fold is recommended as a starting point) and incubation time (2 hours to overnight);

5. IgG cleavage can be easily determined by SDS-PAGE;

6. IdeS protease has a histidine tag, which is easy to remove from the enzyme digestion reaction system;

7. After incubating the digestion product with Protein A beads for 30 to 60 minutes, the purified F(ab ' )2 fragment can be obtained;

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

Frequently asked questions

Q1: If other packaging specifications are required, can it be customized?

A1 ：Yes.