Immunology Exam: Section A - Antibody Engineering and Therapy

This document provides a detailed solution to an Immunology Exam, specifically focusing on Section A. The solution begins by defining and characterizing antibodies, including their binding properties and the role of immunoglobulins. It differentiates between monoclonal and polyclonal antibodies, explaining their production by B-lymphocytes and plasma cells, and the process of creating hybridoma cells for monoclonal antibody production. The document then elaborates on the generation process of monoclonal antibodies, starting with the isolation of splenocytes from mice, immunization with antigens, and the fusion of splenocytes with myeloma cells using polyethylene-glycol. It also discusses the screening and selection of clones based on antigen specificity and immunoglobulin class, followed by functional characterization using ELISA to validate antibody properties. The document further explains the scaling-up process for clones and the therapeutic potential of engineered antibodies, including in vitro hybridoma production and the role of B cells from immunized animals. The solution references several scientific articles to support its explanations, providing a comprehensive understanding of the topics covered in the exam section.

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IMMUNOLOGY EXAM SECTION A
IMMUNOLOGY EXAM – SECTION A
Name:

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IMMUNOLOGY EXAM SECTION A
SECTION A
Solution 1. A.
The main characteristics of antibodies encompass the response to the antigens in our body.
The antibodies are referred to as immunoglobins too. The antibodies have the characteristic to
be bound with the cell membrane and if this does not take place, the antibodies tend to remain
free. The production of antibodies takes place with the Blymphocytes along with the plasma
cells. The mature cells of plasma are being produced which results in the peculiar
characteristic linked to antibodies. The monoclonal antibody is characteristic to the single one
and leads to production of B cell. This antibody only binds to the unique epitope. The
isolation of B-cell is unique in case of monoclonal antibody. The Polyclonal antibody on the
other hand is representative of a greater collection of the B cells. These antibodies well
recognize the epitopes linked to a similar antigen. 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.
Solution 1. B.
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.

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IMMUNOLOGY EXAM SECTION A
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
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.

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IMMUNOLOGY EXAM SECTION A
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
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.
The requirement of 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.

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IMMUNOLOGY EXAM SECTION A
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.
Solution 1 C.
The therapeutic potential, that of engineered antibody is unique. 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 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
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.