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New MAb Description

When you are interested in generating MAbs that meet your specific needs, the ABCF will assist you throughout the entire process.
You can choose any combination of the 8 following services:

Consultation:

We are here for you before, during and after the process of making new MAbs to provide advice enabling you to make informed choices. This is most important when deciding how to:
  • Present the antigen to the animals, because source of your antigen greatly influences the spectrum of MAbs the animal will generate.
  • Identify which hybridoma amongst the thousands generated produces the MAb that works in your assay, because ... What you screen for is ... What you get ...
  • Request a consultation appointment via email here.

Immunization:

  • 5 mice or 4 rats or 4 hamstesr
  • 6 months of housing
  • Up to 5 immunizations
  • Pre-immune sera and up to 4 test seras
Advantage: Choosing this service eliminates the time and hassle involved in getting an IACUC protocol approved.

Fusion:

  • Final boost of an animal (IV or IP challange with the antigen without adjuvant)
  • Fusion of the splenocytes with an appropriate myeloma
  • Plating and selection the fusion products in about 25 – 96 wells plates
  • Qualitative analysis of the how many hybridomas there will be to screen
  • One feeding
Advantage: Although the fusion protocol is relatively simple, highly efficient fusions require some practice.

Maintenance of plated fusion:

  • Provide conditioned media from each well (culture) as it is ready for you to screen for functional MAb
  • Feed the plates
  • Expand positive wells (cultures) and provide more conditioned media for secondary/tertiary screens
Advantage: Opting for this service allows those doing the screening to concentrate on finding the antibodies and not the laborious task of keeping the cells alive.

ELISA screening:

  • Prove that we can perform the ELISA you develop in your lab
  • Perform this triage assay on all wells (cultures) to be screened
  • Analyze the data and identify positive wells (cultures)
Advantage: If the number of lab members available to screen the fusion is limited, it may be helpful to contract the facility to perform the routine ELISA so the people with the greatest degree of experience with the antigen and techniques can focus on screening and finding MAb with the desired functionality after the pass the first filter, the ELISA.

Freezing positive cultures:

  • Freezing a minimum of 10 positive wells, more if you like
  • Transfer as many positive wells to you for freezing (no additional charge)
  • Transfer 1 vial to your lab of the positive culture the facility froze
  • Store 1 duplicate vial in the facility as a back up of the positive culture the facility froze

Cloning or subcloning:

Once a hybridoma culture has been identified that produces a MAb with the sought functionality, the culture NEEDS to be cloned at least once to establish a stable line from which the MAb can be reliably produced.
If you request this service, we will:
  • Plate the culture in semi-solid media
  • Pick all hybridomas that produce MAb into a 96-well, with the ClonePixFL
  • Provide media from these clone for you to determine if the MAb it produces functions in your assay
  • Expand and freeze up to 4 clones that score positive in your assay
  • Transfer 1 vial to your lab of the positive culture the facility froze
  • Store 1 duplicate vial in the facility as a back up of the positive culture the facility froze

Freeze spleen from immunized animals:

  • Final antigen boost of an animal(s)
  • Prepare splenocytes and freeze
  • Transfer all the vials to you
Advantage: If you have other good responding animals, you can have their splenocytes frozen for future fusion, allowing you to keep the repertoire of B cells generated in the animal longer than the animal can survive.

The facility can be reached at (646) 888-2331 for any questions you may have.