Frequently Asked Questions About Mouse Monoclonal Isotyping
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- What is isotyping and subtyping?
- What types of antibodies does this work with?
- What matrices does it work with?
- How sensitive is it?
- What types of samples can I use it in?
- Do I need to dilute my samples?
- Why is it on three separate LFD strips?
- What other Isotypes and Subtypes are there and why do we not provide a test for them?
- What are the phases in the MAb development or production process where isotyping may be used?
- Is isotyping required of all MAbs that are developed or produced? Does the USDA require Isotype classification?
- Who uses isotyping tests?
- What other isotyping kits are available?
- What are the benefits of EnviroLogix Mouse Monoclonal Isotyping kit versus other isotyping tests?
- How are the IsoQuick strips packaged and what is our pricing structure?
- Why don’t the EnviroLogix IsoQuick strips have the interpretation guide printed right on each strip?
- What is isotyping and subtyping?
Isotyping is a classification of monoclonal antibodies based on the characteristics of the Immunoglobulin (Ig) in the sample. There are two formats of isotypes – heavy chain and light chain. The heavy chain includes all the Ig types and subtypes, while light chain includes the formats kappa and lambda.
Immunoglobulin has several characteristics and certain types are more specific than others. IgG (regardless of subtype) is more specific than either IgA or IgM.
- What types of antibodies does this work with?
The IsoQuick strips are designed to be used in the heavy chain and light chain classification of monoclonal antibodies (MAb) made using a mouse host.
- What matrices does it work with?
The IsoQuick strips will work to classify mouse monoclonal antibodies only.
- How sensitive is it?
The sensitivity for each IsoQuick strip is 0.1 µg/mL (ten nanograms per microliter)
- What types of samples can I use it in?
The IsoQuick strips can be used in ascites, cell culture supernatant and purified monoclonal antibodies.
- Do I need to dilute my samples?
If you are using IsoQuick strips in ascities, then you need to dilute your sample to a concentration of 1:10,000 to 1:100,000.
If you are using IsoQuick strips in cell culture supernatant no dilution is required. The strips can be used directly in your sample with a recommended sample size of 150 mL to 500 mL.
If you are using IsoQuick strips in purified MAbs, we recommend you dilute the sample to 0.05-2 mg/mL.
- Why is it on three separate LFD strips?
By dividing the IsoQuick test onto three separate LFD strips, EnviroLogix is providing the user with the greatest flexibility both in their use of the strips and in their purchasing of strips. Because it is more common to classify MAbs at different times in the development or production process, and to classify for different isotypes during the process, the EnviroLogix IsoQuick isotyping strips give the user the freedom to use just what is needed, when it is needed.
- The IgG Strip includes tests for heavy chain isotypes and subtypes IgG1, IgG2a, IgG2b and IgG3.
- The IgA/M strip includes tests for heavy chain isotypes IgA and IgM.
- The K/L strip includes tests for light chain isotypes kappa and lambda.
- What other Isotypes and Subtypes are there and why do we not provide a test for them?
There are several other isotypes and subtypes that may be developed including IgD, IgE and IgG2c. These are rarely used in the development of monoclonal antibodies, and low demand for these tests resulted in their being eliminated from the IsoQuick design specification.
- What are the phases in the MAb development or production process where isotyping may be used?
The process of developing a monoclonal antibody can be separated into three (3) basic phases. Phase 1 is screening. Phase 2 is Purification and Testing. Phase 3 is validating and producing.
Isotyping will be used in Phase 1 screening to help the scientist determine whether the development of the monoclonal antibody is on target with the isotype required by the specification. This will usually entail isotype testing for heavy chain types, with IgG subtypes being the mostly widely sought isotype classifications.
In Phase 2 testing will be more specific for the isotype or subtype that was specified in the development plan and that was identified in Phase 1. A change or inconsistency in isotype classification in phase 2 may mean restarting the development process or retracing steps to a known point earlier in the development cycle.
In Phase 3 testing will be done to confirm the heavy chain classification of the MAb, and to determine the light chain type as well. Light chain typing may be performed in either Phase 1 or Phase 2, but it is most often conducted as a confirmation step in Phase 3.
In the case of a new MAb that is developed both the heavy chain and light chain characteristics will be identified as part of the finished product specifications.
- Is isotyping required of all MAbs that are developed or produced? Does the USDA require Isotype classification?
The USDA and other regulatory bodies may require isotype classification for certain monoclonal antibodies developed for human diagnostics or human treatments. It is not clear if other agencies require isotype classification for monoclonal antibodies not directly applied to human health care.
- Who uses isotyping tests?
Any company engaged in the development, purification or production of monoclonal antibodies, including in-house labs and outsourced development or production facilities will use Isotyping tools. There are thousands of labs engaged in the development, purification and production of monoclonal antibodies. A large percentage, but not all labs developing monoclonal antibodies are using a mouse as a host.
- What other isotyping kits are available?
There are a number of LFD strip format isotype kits available including a two strip kit from Roche, a two strip kit from MicraTech and a single strip kit from BioAssay Works. The Roche kit offers the same scope as IsoQuick – that is it offers classification for the IgG subtypes, IgA, IgM and kappa lambda. The two strip kit from MicraTech offers only the IgG subtypes and IgA and IgM isotypes. It does not offer the light chain kappa/lambda classification. The Bioassay Works Isostrip kit is a single strip test contained in a plastic holder (like a pregnancy test), and it too provides classification for IgG and subtypes, as well as for IgA and IgM. It does not provide kappa/lambda classification.
In addition, there are a number of ELISA plate isotyping tests. These may offer all isotype classifications, and are much less costly to run per test. These isotype tool formats will only be used by very large labs producing hundreds or thousands of monoclonal antibodies annually.
- What are the benefits of EnviroLogix Mouse Monoclonal Isotyping kit versus other isotyping tests?
The IsoQuick Kit for Mouse Monoclonal Isotyping offers several advantages over other products in the market.
- Flexibility – Dividing the core tests onto three LFD strips, EnviroLogix is giving the user the greatest flexibility in purchasing and using.
- Convenience – the IsoQuick strips are provided in simple desiccated tubes in packages of five (5) and twenty-five (25) tests.
- Speed – IsoQuick tests develop positive test lines in five minutes.
- Ease of Use – IsoQuick strips can be used “neat” in cell supernatant and purified MAbs. Dilution is not required, and testing can be done directly in the sample vessel. Some dilution may be required for testing in ascities, however, the same ease of use applies once the test sample is prepared – insert the IsoQuick strip and wait five minutes.
- Accuracy – Each IsoQuick strip meets or exceeds the sensitivity specifications of other LFD isotyping products on the market including Roche strip. The ELISA plate tests may or may not be more sensitive.
- Shelf-life – The IsoQuick strips will provide a minimum of twelve (12) months useful shelf-life. Users need not worry about over-purchasing and having tests go bad on the shelf.
- How are the IsoQuick strips packaged and what is our pricing structure?
EnviroLogix IsoQuick Mouse Monoclonal Isotyping strips will be sold in packages of five (5) strips and twenty-five (25) strips. You may purchase one or more of each of the strips in either canister size. A kit including one five-strip canister of each of the three IsoQuick strips is also available.
Please call for specific pricing information, including volume discounts.
- Why don’t the EnviroLogix IsoQuick strips have the interpretation guide printed right on each strip?
Due to the need for specialized wide LFD production equipment and specialized printing equipment, EnviroLogix is not able to print the identity of each result line directly on the strip. EnviroLogix provides two results interpretation tools that are convenient, easy to use and that will not blur or contaminate the test line. Each IsoQuick canister will have a life size interpretation guide on the side. Additionally, each kit ordered will include a laminated interpretation and user guide for all three strips.
PDF files, where provided, require Adobe Acrobat Reader.
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