Biomedical Engineering Reference
In-Depth Information
not produce any of the antibody molecule which is required for producing
cells. For the production of antibody a fusion of myeloma cell with the cell
shaving ability of growth. Mice when immunised with the antigen produces
polyclonal antibodies. Spleen cells are fused with non-antibody producing
myeloma cells using polyethylene glycol by removing the spleen of mouse
and as it contain a very high concentration of B lymphocytes. h e mem-
branes of both cells fuse and merge. h e cell nucleus merges, then the chro-
mosomes are mixed and the immortalised antibody producing hybridoma
cells are obtained from it. Unfused spleen cells die and the myeloma cells
which are unfused are terminated on a selective HAT medium. h e result-
ing mixture of polyclonal hybridoma is grown in a culture in a well manner,
divided and is diluted. If the supernatant part of the well contains specii c
polyclonal antibodies, the well is further diluted, divided and synchronised
until the hybridomas in the well are cloned from only a single parent cell,
producing one type of antibody only. Later, a monoclonal antibody pro-
ducing hybridoma is derived from it. Normally, this procedure yields many
polyclonal hybridomas, which produce specii c antibodies. h e monoclonal
antibodies produced are tested for ai nity, sensitivity and specii city and a
few hybridoma cells are chosen for further development of the cells.
Monoclonal antibodies can be grown, in culture l asks or bioreactors.
Monoclonal antibodies can also be grown in vivo under composite methods.
Hybridoma cell are injected into the peritoneum of mice in a very decent
manner and the tumour-like growth rate also produces large amounts of
antibodies in ascetic l uid with the help of innoculation. In vitro methods
delivers pure antibody which has low yields and the in vivo method results in
high yields of antibody, which are contaminated and specii ed with proteins
and other antibodies, and purii cation is very necessary.
7.8.1.7
Antibody fragments and Recombinant Antibodies
A genetic technique namely called as combinatorial phage display allows
for the production of the Fab fragment of antibodies, by combining and
attributing the genes for these specii c regions with phage particles and
consequently producing the Fab fragment of antibody in bacteria cells
[143]. h e two Fab fragments of an antibody are identical, which contain
two identical binding sites; it is also possible to create antibodies with two
dif erent binding sites.
h e bi-functional antibodies can be created chemically by cleaving and
forming the disulphide bonds and cross linking one of the bi-functional
antibodies with another antibody fragment. Biological production is
achieved by fusing two hybridoma cells the resultant antibody is generally
