In monoclonal antibody production, monoclonal cells are differentiated from?
ANSWER: (C) Myeloma cell linesREF:Applications and engineering of monoclonal antibodies by David J. King page 17, A practical guide to monoclonal antibodies by J. Eryl Liddell, Anthony Cryer Page 69"A myeloma cell is classically defined as a neoplastic plasma cell in myelomatosis producing a structurally and electrophoretically homogeneous (monoclonal) paraprotein"Fusion of the myeloma cells and the harvested B cells is usually accomplished using polyethylene glycol (PEG) as an agent to induce membrane fusion (cheep, reliable and most commonly used method). Alternatively electrofusion or Sendai virus can be used. After fusion a mixture of spleen cells myeloma cells and hybridoma cells are present and next step is selection of hybridoma cells over other cellsPOST FUSION SELECTION:Myeloma cells are deficient in HGPRT and are not able to use salvage pathway for RNA synthesis. HAT (Hypoxanthine, Aminopterin, Thymidine) medium is used to achieve selection. Aminopterin is an inhibitor of DNA and RNA synthesis and thus blocks myeloma cells. Hybridoma cells however have HGPRT and thus can add hypoxanthine and thymidine to produce RNA and thus can survive.Cell typeDNA synthesisSurvival in HAT medium Salvage pathwayDe Novo pathway MyelomaHGPRT-Aminopterin sensitiveDie (no DNA symthesis)SpleenHGPRT +Aminopterin sensitiveDie (finite survival in vitro)Myeloma spleen hybridHGPRT +Aminopterin sensitiveLiveMyeloma myeloma hybridHGPRT-Aminopterin sensitiveDie (no DNA symthesis)Spleen spleen hybridHGPRT +Aminopterin sensitiveDie (finite survival in vitro)Alternatively myeloma cells can be selected to be deficient in enzyme thymidine kinase (TK) by growth in bromodeoxyuridine.
Sensitized B cells
Sensitized T cells
Myeloma cell lines
None of these
Miscellaneous (Structure & Function of Proteins)