Monoclonal Abs - 4
Recombinant
DNA technique:
emRNA is
isolated from lymphoid tissue of donor who has been
sensitized with a specific antigen
eisolated mRNA
is converted to dsDNA using reverse transcriptase + DNA polymerase
enzymes
epolymerase
chain reaction (PCR) is used to amplify the gene fragments of VH and VL
ethe amplified
fragments of VH and VL are then injected into E. coli
bacteriophage via a plasmid vector and gene fragments fuse
with bacteriophage protein
eeach
recombinant bacteriophage contains a different combination of
VH and VL genes thus producing antibodies of differing
specificities.
ethese
specificities are then identified by reacting with different
antigens and are then cataloged into a monoclonal antibody
library
