HAT medium
B-cell
.HAT Medium (genes that encode them.
The enzyme
tetrahydrofolate (THF) by the reduction of dihydrofolate, is specifically blocked by aminopterin. THF, acting in association with specific proteins
, can receive single carbon units that are then transferred to specific targets.
One of the important targets for cellular reproduction is
DNA polymerases to create DNA. Without the THF required to convert dUMP, there can be no TTP, and DNA synthesis cannot proceed, unless TMP can be produced from another source. The alternative source is the thymidine present in the HAT medium that can be absorbed by the cells and phosphorylated by thymidine kinase
(TK) into TMP.
The synthesis of IMP, (precursor to GMP and GTP, and to AMP and ATP) also requires THF, and also can be bypassed. In this case
PRPP, liberating pyrophosphate
, to produce IMP by a salvage pathway.
Therefore, the use of HAT medium for cell culture is a form of
revertant colonies.[1]
Applications
HAT medium is used for preparation of
antibodies
(a property of B cells) and are immortal (a property of myeloma cells).
The incubated medium is then diluted into multiwell plates to such an extent that each well contains only 1 cell.
Then the monoclonal antibodies
.
The production of monoclonal antibodies was first invented by César Milstein and Georges J. F. Köhler, which earned them the 1984 Nobel Prize in Physiology or Medicine, shared with Niels Kaj Jerne.
References
- PMID 9671746.