Detailed Explanation of Monoclonal Antibody Production in Immunology

This document provides a detailed explanation of monoclonal antibody production. The process begins with the generation of hybridoma cells through the fusion of desired antibody-producing splenocytes and myeloma cells, often sourced from mice. The document outlines the screening and selection of clones based on antigen specificity and immunoglobulin class, followed by functional characterization and validation using techniques like ELISA. The scaling-up process involves expanding clones to produce antibodies in larger volumes, often using bioreactors. The production time is significant, recognizing a single epitope based on the antigen. The in vitro production method involves immunizing animals with a specific antigen, removing B cells from the spleen, and fusing them with immortal cancerous cells (myeloma cells) to yield hybridoma cells. These cells are then placed in a selective medium that allows continuous growth, essential for producing large quantities of monoclonal antibodies. The document also references several research papers to support the information provided.

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IMMUNOLOGY EXAM SECTION A
IMMUNOLOGY EXAM – SECTION A
Name:

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IMMUNOLOGY EXAM SECTION A
SECTION A
Q 1 (b) . Discuss the production of Monoclonal Antibodies
Solution
The monoclonal antibody does start the generation process with the cells produced, that of
the monoclonal antibody producing cells (Dunand et al., 2016, pp. 812; Han et al., 2018, pp.
30; van de Donk, 2016, pp. 691). These cells are referred to as hybridomas and can be
obtained from the fusion taking place of desired antibody producing splenocytes along with
myeloma cells.
The cells are naturally being sourced from the animals. The most generic form of source
considering animals is the mice (Dunand et al., 2016, pp. 812; Han et al., 2018, pp. 30; van de
Donk, 2016, pp. 691). The cell fusion along with the larger number of clones are able to get
screened and also being selected with the basis of antigen and this is the class pertaining to
the immunoglobin and the antigen (Ecker et al., 2015, pp. 11; Swanstrom et al., 2016, pp.
1124; van de Donk et al., 2016, pp. 694).
The process starts with the lining up of the cell lies, that are hybridoma and are uniquely
being identified with each hit being well confirmed. The validation of such a hit has also to
take place. The characteristic that is persistent with a variety of functional assays of
downstream nature are significant (Dunand et al., 2016, pp. 812; Han et al., 2018, pp. 30; van
de Donk, 2016, pp. 691). The hybridoma cell lines can well be identified and the completion

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IMMUNOLOGY EXAM SECTION A
of this process leads to the next stage. This stage is of the clones being scaled up wherein
additional downstream bioprocesses do takes place.
The process of production of monoclonal antibodies takes place with the inception of mice
and isolation with respect to the splenocytes. The mice are being immunized with the antigen
and the process leads to the stage wherein the blood gets screened for the production of
antigens.
The splenocytes that are anti-body producing are being isolated for the vitro hybridoma
production. The proceeding stage is associated with the preparation, that of the Myeloma
cells (Ecker et al., 2015, pp. 11; Swanstrom et al., 2016, pp. 1124; van de Donk et al., 2016,
pp. 694). The myeloma cells are well immortalized and tend to be fused with the spleen cells.
The resultant of such a fusion taking place is the hybridoma capabilities with the growth that
is of unlimited nature.
The fusion process gets ready with the myeloma cells. The fusion process tends to be crucial.
The fusion is remarked with the isolation of splenocytes being fused in order to form
hybridomas. The process takes place in the presence of the polyethylene-glycol. The process
leads to the cell membrane being fused (Dunand et al., 2016, pp. 812; Han et al., 2018, pp.
30; van de Donk, 2016, pp. 691). The clone screening is well selected on the basis of antigen
and the process involves specificity along with the immunoglobin class.
There tend to be functional characterization in order to confirm and also validate the
characteristics. ELISA is linked to the confirmation and such a validity. The potential of such
functional characteristics leads to the higher producing quality. The scaling up process

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follows this. This stage is marked with the. Scaling up done for the clones producing the
desired antibodies along with the wean with respect of the selection agents (Ecker et al.,
2015, pp. 11; Swanstrom et al., 2016, pp. 1124; van de Donk et al., 2016, pp. 694). The
expansion process includes the expansion taking place with respect to the clones. This aids in
the production of antibodies in a greater volume. The bioreactors or the flasks can be
provisioned as an example.
He assays do require the refined antibody along with the affinity which is obtained utilizing
the polyclonal antiserum (Dunand et al., 2016, pp. 812; Han et al., 2018, pp. 30; van de Donk,
2016, pp. 691). The production time taken for the monoclonal antibodies is significantly
more. The production time is increased with the process taking lot of sub processes into
account.
The larger quantities linked to the production recognizes the single epitome which is based,
that of the antigen. The production tends to take place with the continuity and uniformity
wherein the hybridoma is being made. The production of monoclonal antibodies is being
done considering the production in vitro (Dunand et al., 2016, pp. 812; Han et al., 2018, pp.
30; van de Donk, 2016, pp. 691). This is being well utilized with the technique asking to
tissue culture. The monoclonal antibodies get produced in a manner that the immunization of
the animals is significantly proceeded with the specific antigen (Ecker et al., 2015, pp. 11;
Swanstrom et al., 2016, pp. 1124; van de Donk et al., 2016, pp. 694). The role of the B cells
too originating from the spleen with respect to the immunized animal is proceeded with the
removal of the same.

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IMMUNOLOGY EXAM SECTION A
The normal cells do not have the capacity to proliferate forever. In this case, the fusion needs
to take place. The same is being done with the immortal cancerous cells which are referred to
as the myeloma cells and the process leads to the yield of hybridoma cells (Dunand et al.,
2016, pp. 812; Han et al., 2018, pp. 30; van de Donk, 2016, pp. 691). The cells are being
placed in the medium which is of selective nature and does tend to allow the growing up of
cells continuously (Ecker et al., 2015, pp. 11; Swanstrom et al., 2016, pp. 1124; van de Donk
et al., 2016, pp. 694). The process is crucial as the larger quantities of production, that of the
Monoclonal Antibodies takes place.

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References
Dunand, C.J.H., Leon, P.E., Huang, M., Choi, A., Chromikova, V., Ho, I.Y., Tan, G.S., Cruz,
J., Hirsh, A., Zheng, N.Y. and Mullarkey, C.E., 2016. Both neutralizing and non-neutralizing
human H7N9 influenza vaccine-induced monoclonal antibodies confer protection. Cell host
& microbe, 19(6), pp.800-813.
Ecker, D.M., Jones, S.D. and Levine, H.L., 2015, January. The therapeutic monoclonal
antibody market. In MAbs (Vol. 7, No. 1, pp. 9-14). Taylor & Francis.
Han, M., Rock, B.M., Pearson, J.T. and Rock, D.A., 2016. Intact mass analysis of
monoclonal antibodies by capillary electrophoresis—Mass spectrometry. Journal of
Chromatography B, 1011, pp.24-32.
Swanstrom, J.A., Plante, J.A., Plante, K.S., Young, E.F., McGowan, E., Gallichotte, E.N.,
Widman, D.G., Heise, M.T., De Silva, A.M. and Baric, R.S., 2016. Dengue virus envelope
dimer epitope monoclonal antibodies isolated from dengue patients are protective against
Zika virus. MBio, 7(4), pp.e01123-16.
van de Donk, N.W., Moreau, P., Plesner, T., Palumbo, A., Gay, F., Laubach, J.P., Malavasi,
F., Avet-Loiseau, H., Mateos, M.V., Sonneveld, P. and Lokhorst, H.M., 2016. Clinical
efficacy and management of monoclonal antibodies targeting CD38 and SLAMF7 in multiple
myeloma. Blood, 127(6), pp.681-695.