No two research projects are exactly alike but it is important that the processes and procedures you use are predictable and replicable. Inotiv has more than 20 years of experience refining our approach to developing monoclonal antibodies from a wide variety of peptides and proteins. While our procedures are well-defined, we are flexible when we need to be.
When you partner with us, our scientists will work with you to develop a procedure that will best meet your needs, with processes that are consistent and service that is responsive.
Our production process is broken down into three stages which allows you to pay for only those stages that are completed. Once the desired clones are developed, we will scale up some material for your in vivo or in vitro use.
Our schedule for producing up to three positive clones includes:
STAGE I: Immunization
Ten virus antibody-free mice will be immunized with the customer's antigen. The mice will be housed in an isolator during the immunization period. We immunize ten mice to increase our chance of an immunogenic reaction
In order to get a better antibody response, the antigen is prepared with an adjuvant such as FCA (Freund's complete adjuvant) for the primary immunization
The emulsion contains heat-killed mycobacterium that ensures the antigen is released slowly into the animal's circulatory system
The bacteria in the adjuvant stimulates the animal’s immune system
Additional boosts or immunizations are necessary for high antibody levels
FIA (Freund's incomplete adjuvant) is used
Common routes for injection are i.p., i.m., i.v., s.c.
For hybridoma development, s.q. is typically used. five-to-100 ug of antigen are used for each immunization. We ask the client for two-three mg of antigen
At six weeks, following the primary immunization and the first boost, the mice will be tested for antibody titer. The antibody will be determined by ELISA assay unless the customer has an alternative assay.
If additional boosts are required, they will be administered and the mice will be retested for antibody titer
Once immunization is complete, the mice will be held for three-to-four weeks before fusion
Three-to-five days before the fusion, a mouse will be selected for the pre-fusion boost
2) Screen and select
1) First cloning
2) Final cloning
3) Selection or non-HT requirement
4) Frozen stock
NOTE: Our service allows for the development of up to three, five or ten positive clones.
Discover how Inotiv helps companies in providing custom antibody services, click here to read more.
Get in touch — Contact our antibodies and bioproducts team
At Inotiv, we are dedicated to helping you secure the potential of your product by providing you with a comprehensive selection of research animals. Our experts are ready to assist you.
If you have a more general inquiry click here to get in touch — general inquiry form.