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Producing monoclonal antibodies

Some types of assays require better antibody specificity and affinity than can be obtained using a polyclonal antiserum. To attain this high specificity, all of the antibodies must bind with high affinity to a single epitope. This high specificity can be provided by monoclonal antibodies (mAbs) . [link] compares some of the important characteristics of monoclonal and polyclonal antibodies.

Unlike polyclonal antibodies, which are produced in live animals, monoclonal antibodies are produced in vitro using tissue-culture techniques. mAbs are produced by immunizing an animal, often a mouse, multiple times with a specific antigen. B cells from the spleen of the immunized animal are then removed. Since normal B cells are unable to proliferate forever, they are fused with immortal, cancerous B cells called myeloma cells, to yield hybridoma cells. All of the cells are then placed in a selective medium that allows only the hybridomas to grow; unfused myeloma cells cannot grow, and any unfused B cells die off. The hybridomas, which are capable of growing continuously in culture while producing antibodies, are then screened for the desired mAb. Those producing the desired mAb are grown in tissue culture; the culture medium is harvested periodically and mAbs are purified from the medium. This is a very expensive and time-consuming process. It may take weeks of culturing and many liters of media to provide enough mAbs for an experiment or to treat a single patient. mAbs are expensive ( [link] ).

Diagram showing production of monoclonal antibodies. Antigen is injected into an animal (such as a mouse) Spleen cells are extracted. Myeloma line cells from a cell culture are added to the spleen cells in a test tube. Then, hybrid cells are selected and grown. Hybrid cells are separated and allowed to proliferate into clones (hybridomas). Each hybrid produces a different antibody and the desired antibody is selected. This hybridoma is then grown to produce large batches of desired mAB.
Monoclonal antibodies (mAbs) are produced by introducing an antigen to a mouse and then fusing polyclonal B cells from the mouse’s spleen to myeloma cells. The resulting hybridoma cells are cultured and continue to produce antibodies to the antigen. Hybridomas producing the desired mAb are then grown in large numbers on a selective medium that is periodically harvested to obtain the desired mAbs.
Characteristics of Polyclonal and Monoclonal Antibodies
Monoclonal Antibodies Polyclonal Antibodies
Expensive production Inexpensive production
Long production time Rapid production
Large quantities of specific antibodies Large quantities of nonspecific antibodies
Recognize a single epitope on an antigen Recognize multiple epitopes on an antigen
Production is continuous and uniform once the hybridoma is made Different batches vary in composition

Clinical uses of monoclonal antibodies

Since the most common methods for producing monoclonal antibodies use mouse cells, it is necessary to create humanized monoclonal antibodies for human clinical use. Mouse antibodies cannot be injected repeatedly into humans, because the immune system will recognize them as being foreign and will respond to them with neutralizing antibodies. This problem can be minimized by genetically engineering the antibody in the mouse B cell. The variable regions of the mouse light and heavy chain genes are ligated to human constant regions, and the chimeric gene is then transferred into a host cell. This allows production of a mAb that is mostly “human” with only the antigen-binding site being of mouse origin.

Questions & Answers

what is the size of virus
Beatrice Reply
What is the difference between TVC and Bioburden test
Mohamed
?
Mohamed
structure of bacteria and 10 types
Jennifer Reply
what is accidental host?
Domingo Reply
what is endomembrane system
Ikpi Reply
what is human anatomy
Ikpi
okay. Go ahead and ask
Blessing Reply
Industrial microbiology mcq
mohamed
Okay. What's your question?
Blessing
life arises from living matter or live organism.
Swami Reply
yes
sildra
I think live matter arises from non living matter
sildra
I dont think so...can u explain with an example
Manya
living maters made by non living matters
sildra
non living matters like stones? rocks?
Manya
eno
sildra
no
sildra
then?
Manya
cells are made by C N O minerals etc
sildra
I mentioned these as non living maters
sildra
that's all
sildra
cells are made up of those things but they originate from living things..
Manya
ok
sildra
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Manya
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sildra
thanks
sildra
India...u?
Manya
Tamil nadu
sildra
I am from Maharashtra
Manya
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sildra
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Manya
same
sildra
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sildra
yes i am
Manya
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sildra
what s the scope for micro in ur state?
Manya
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sildra
have to give an entrance exam for every college here...so lets c
Manya
food industries, medical lab, vaccine industries ,etc
sildra
hoping for pune University...wbu?
Manya
great!
Manya
is that centeral University right
sildra
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sildra
hello
Udhaya
Hi
Maruf
Family kindly help me with this question? 1) Shortlist the configurative measurements of the following human anatomical ranges of÷ - Blood ( haemeglobin) in both male and female - Haematocytes in both male and female - Hepatocytes in both male and female - Lymphocyte / T. Lymphocytes in both male
Gifted
My names are Gift Mwale and am a Zambian. Kindly help me with this research which goes like this... 1) Shortlist the configurative measurements of the following human anatomical ranges of ÷ - Blood ( haemeglobin) in both male and female - Hepatocytes in both male and female - Haematocytes in both
Gifted
please what is the full meaning for TCDS
UDEME
from a single cell
Freedom
tcds means transcranial direct current stimulation...in this small electric currents are given to brain( specific parts) to help increase brain performance or to help with depression.. current should be in range 0.5-2.0mA
Manya
what's underlying disease relating unsanitary diet microorganism with the highest rate of epidemology solution and efficacy leading molecules elucidated structural solutions
feven
please can anybody talk about brain tumour and its cure.
BELLO Reply
enlargement of the thyroid gland resulting in over production of hormone.
Kamal Reply
What can u say on Thyroid Cancer?
Abdulkareem
Please, talk about the thyroid cancer.
BELLO
explain the Grave's disease
John Reply
what is cell
Avi Reply
is unit of life
Kamaluddeen
Ok
mohamed
who is an industrial microbiologist
Cynthia Reply
I want to know the biochemical composition of bacteria
Josh Reply
It contains peptidoglcon, DNA nd RNA
Asiya
what are Carrier protein
Ikpi
bacteriophage disadvantage
Momina Reply
disease due to __________ abnormalities are termed primary immunodeficiencies
Tayee Reply
Some primary immunodeficiencies are due to a defect of a single cellular or humoral component of the immune system.
Prince
Examples of primary immunodeficiencies include: chronic granulomatous disease, X-linked agammaglobulinemia, selective IgA deficiency etc
Prince
thank you
Nana
explain microbial mutation
Emerald
what is mutation
Cynthia Reply
alteration in genetic makeup
UDEME

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Source:  OpenStax, Microbiology. OpenStax CNX. Nov 01, 2016 Download for free at http://cnx.org/content/col12087/1.4
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