Hi All
I recently have got some hybridoma to generate mouse mAb to a antigen of interest. However we found the mAbs produced are IgG1, while we would like to have IgG2a. Is there anyway to work from these hybridoma and make them to produce IgG2a rather than IgG1?
Selecting for low frequency class-switch variants of hybridomas is generally quite a laborious process. You need need to have a selection process that allows you to detect the switched hybridomas and then subclone them e.g. flow cytometry based sorting. Or alternatively you can carry out cell cloning and try to detect which colonies are making the appropriate isotype. This might entail several rounds of enrichment of colonies seeded at multiple cells per well to enable you to screen enough cells to pick the rare class-switched cells (which might be present at frequencies of around 1 in a million or less). See for example Hale,G.; Cobbold,S.P.; Waldmann,H.; Easter,G.; Matejtschuk,P.; & Coombs,R.R.A. Isolation of low-frequency class-switch variants from rat hybrid myelomas J. Immunol. Meth. 103, 59-67 (1987)
However an alternative approach that is probably much more amenable is simply to sequence or clone the V-regions from your hybridoma and then express a recombinant version formulated exactly how you'd like it. I can recommend a company that would do that for you if you wished Absolute Antibody Ltd http://absoluteantibody.com/
As a declaration of interest I'm associated with Absolute Antibody Ltd.
http://absoluteantibody.com/about-us/
try also:
Isolation of immunoglobulin class switch variants from hybridoma lines secreting anti-idiotope antibodies by sequential sublining.
Müller CE, Rajewsky K.
J Immunol. 1983 Aug;131(2):877-81.
PMID: 6190943 [PubMed - indexed for MEDLINE]
190.
Isolation of heavy chain class switch variants of a monoclonal anti-DC1 hybridoma cell line: effective conversion of noncytotoxic IgG1 antibodies to cytotoxic IgG2 antibodies.
Parham P, Kipps TJ, Ward FE, Herzenberg LA.
Hum Immunol. 1983 Oct;8(2):141-51.
PMID: 6417068 [PubMed - indexed for MEDLINE]
I suspect there are quite a few more
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology
- Virology
- Virus