Production and Purification of Recombinant Antibodies from Mammalian Cells
Transcrição
Begin with a tube containing separate vectors for the heavy-chain and light-chain antibody genes.
Add a transfection reagent to form complexes with the vectors.
Transfer this mix to mammalian host cells and incubate for internalization.
Add an anti-clumping agent to promote the even distribution of transfected cells, improving cell recovery and growth.
Add an anti-bacterial-anti-mycotic solution to prevent contamination.
Within the host cell, the vectors integrate into the host genome.
Following gene transcription, individual mRNAs exit the nucleus and undergo translation to form nascent antibody chains.
Proper folding and modification of the nascent chains yields heavy and light chains.
These chains combine to form fully functional antibodies, which are then released into the extracellular medium.
Harvest the medium and centrifuge. Filter the supernatant.
Pass the filtrate through a purification column that allows selective binding of the antibodies.
Using an elution buffer, elute the purified antibodies.
