Monoclonal

Antibody
Submitted To:
Md. Hasanuzzaman Shohag

Md. Akash Molla (ID: 1821467049)

Course: PHR325

Section: 03
 Monoclonal Antibody: Production, Application & Limitations

Monoclonal Antibody (abbreviated as mAb or moAb) is a kind of antibody created by cloning a

specific type of white blood cell. A single parent cell may be found in all future antibodies made

in this way. Monoclonal antibodies can only attach to a single epitope if they have monovalent

affinity.  Those antibodies that are identical to that of antibody, which targets one specific

antigen. Single clone of cells.These are also called as identical immunoglobulins.

Monoclonal Antibody in Biotechnology

The development of monoclonal antibodies during the latter quarter of the 20th century was a

major advance in biotechnology. A monoclonal antibody is one that was created through genetic

engineering and other similar methods, such as recombinant DNA techniques.

Origin of Monoclonal Antibody

In order to mass-produce antiviral antibodies, a white blood cell exposed to a particular protein

family is cloned.The name “Monoclonal Antibody”came from: - "Hybridoma" are cells that have

the ability to proliferate indefinitely but generate a particular antibody. Patients with

malignancies that express the human epidermal growth factor receptor 2 (HER-2) protein may

benefit from naked monoclonal antibodies, such as alemtuzumab (Campath, Genzyme) and

trastuzumab (Herceptin, Genentech).

The First Monoclonal Antibody: The FDA approved Orthoclone OKT3 (muromonab-CD3) in

1986 to prevent kidney transplant rejection. This IgG2a monoclonal mouse antibody recognizes

CD3. This molecule binds and blocks T-lymphocyte CD3.

Monoclonal Antibodies Trigger the Immune System: -Outer membranes of cancer cells may be

destroyed by certain monoclonal antibodies. Stopping the growth of new cells. Some antibodies

stop cancer cells from connecting with proteins that help them grow and survive.

Monoclonal Antibodies came from humans: - Antibody discovery may be accomplished using a

variety of methods, but those that have been "humanized" have shown to be the most effective.

>The immune system employs these antibodies to recognize and neutralize foreign things such

as viruses or bacteria.

These antibodies have two major functions:

1. Induce Immune Responses after Binding: -Biochemistry, molecular biology, and medicine all

benefit from the ability to make antibodies. Monoclonal antibodies may be made to recognize

practically any particular chemical. They may then be used to identify or purify the material they

have bound to with the help of these antibodies.

2. Recognize & BindAntigen: -Monoclonal antibodies can only attach to a single epitope if they

have monovalent affinity (the part of an antigen that is recognized by the antibody). Polyclonal

antibodies bind to several epitopes and are produced by a variety of antibody-secreting plasma

cell subtypes.

Production of monoclonal antibody:A single clone of B cell produces a sequence of identical

antibodies known as monoclonal antibodies. A vital tool for biological research and therapeutic

usage, they have been employed extensively since their discovery in the 1980s. Hybridoma

technology is the name given to this process by researchers at Stanford University.G. Smith

developed phage display in 1985 as a second way to make monoclonal antibodies. Large
numbers of peptides, proteins, and antibodies may now be produced using this method.A

bespoke monoclonal antibody service, as well as large-scale antibody synthesis and purification,

are both available from Sino Biological. Regardless of your requirements, Sino Biological can

come up with personalized solutions, and its knowledge will help you throughout the whole

process.A few kinds of tests require preferred counter acting agent explicitness and partiality

over can be gotten utilizing a polyclonal antiserum. To accomplish this high particularity, each of

the antibodies should tie with high fondness to a solitary epitope. This high particularity can be

given by monoclonal antibodies (mAbs).

Dissimilar to polyclonal antibodies, which are delivered in live creatures, monoclonal antibodies

are delivered in vitro utilizing tissue-culture strategies. mAbs are delivered by vaccinating a

creature, regularly a mouse, on various occasions with a particular antigen. B cells from the

spleen of the vaccinated creature are then taken out. Because normal B cells cannot proliferate

indefinitely, they are joined with malignant B cells termed myeloma cells to form hybridomas.

Unfused myeloma cells and unfused B cells are killed when the cells are put in a specific

medium that only permits Hybridomas to develop.The Hybridomas, which are equipped for

filling ceaselessly in culture while delivering antibodies, are then evaluated for the ideal mAbs.

Those delivering the ideal mAbs are filled in tissue culture; the way of life medium is reaped

occasionally and mAbs are cleansed from the medium. This is a pricey and tedious cycle. It

might require a long time of refined and numerous liters of media to give enough mAbs to an

analysis or to treat a solitary patient. mAbs are costly. 

Applications

Diagnostics Test

Once a substance's monoclonal antibodies have been created, they may be used to test for its

existence. In order to identify proteins, Western blotting and immunodot blotting are used. It is

possible to identify antigens using monoclonal antibodies in immunohistochemistry and

immunofluorescence to detect a material with either paraffin - embedded slices or living cells.

Autoimmune Diseases: -There are many monoclonal antibodies used to treat autoimmune

illnesses. Inhibiting IL-2 on activated T cells with basiliximab and daclizumab reduces the risk
of acute rejection after kidney transplantation. For those with mild to moderate allergic asthma,

omalizumab may be an effective treatment.

Analytic and Chemical Uses: -The immunoprecipitation technique may be used to extract target

molecules from mixtures utilizing antibodies.

Therapeutic Uses: -T Antibody monoclonal treatment

Various techniques are used by therapeutic monoclonal antibodies, including inhibiting the

activities of targeted molecules, triggering cell death when the target is expressed, or altering

signaling pathways in those cells that produce the target.

Major Applications of Monoclonal Antibodies are as

1. Cancer therapy (Immuno-toxicology)

To deliver a poison, radioisotope, cytokine, or any active compound, monoclonal

antibodies (mAbs) may be changed. In addition to binding to a target antigen, antibodies

may also attach to a conjugate or effector cell via their Fab regions. The Fc region of

every intact antibody binds to cell receptors and other proteins.Antibody-directed enzyme

prodrug treatment (ADEPT) and antibody-dependent cell-mediated cytotoxicity (ADCC)

are used interchangeably in this article. ADCC is a cancer immunotherapy using

monoclonal antibodies. The FDA for the treatment of cancer has authorized the following

monoclonal antibodies:

 Alemtuzumab

 Bevacizumab

 Cetuximab
  Antineoplastic chemotherapy drug Gemtuzumab-ozogamicin

 Ipilimumab

     2. Radioimmunoassay

     3. Protein Purification

     4. Diagnosis of disease

     5. Gene Cloning and expression

Limitations:                                                                                                                                     

1) Tedious task - anywhere between a half year - 9 months. 

2) Pricey and needs impressive work to deliver them. 

3) Little peptide and piece antigens may not be acceptable antigens-monoclonal immunizer may

not perceive the first antigen. 

4) Hybridoma culture might be dependent upon pollution. 

5) Framework is just very much created for mouse and rodent and not intended for different

creatures. 

6) Over close to 100% of the cells do not make due during the combination interaction –

diminishing the scope of helpful antibodies that can be delivered against an antigen lastly.

Unlimited production source, highly repeatable and scalable. Monoclonal antibodies may be

made indefinitely using Hybridomas as a source. The antibody's quality is consistent throughout
manufacturing batches. No need to worry about caring for the animals when you have the ability

to create antibodies when you need them.On the other hand,only mice and rats benefit from the

system's current state of development; other species do not. An undertaking that will take

between six and nine months to complete 99 percent of the cells die after fusion, which reduces

the available pool of effective antibodies to an antigen.

References:

1. Rodrigues ME, Costa AR, Henriques M, Azeredo J, Oliveira R. Technological progresses in

monoclonal antibody production systems. Biotechnology progress. 2010 Mar;26(2):332-51.

2. Sithigorngul P, Chauychuwong P, Sithigorngul W, Longyant S, Chaivisuthangkura P.

Development of a monoclonal antibody specific to yellow head virus (YHV) from Penaeus

monodon. Diseases of aquatic organisms. 2000 Aug 10;42(1):27-34.

3. Cho YJ, Lee DH, Kim DO, Min WK, Bong KT, Lee GG, Seo JH. Production of a monoclonal

antibody against ochratoxin A and its application to immunochromatographic assay. Journal of

Agricultural and Food Chemistry. 2005 Nov 2;53(22):8447-51.

4. Epenetos AA, Snook D, Durbin H, Johnson PM, Taylor-Papadimitriou J. Limitations of

radiolabeled monoclonal antibodies for localization of human neoplasms. Cancer research.

1986 Jun 1;46(6):3183-91.

5. Parray HA, Shukla S, Samal S, Shrivastava T, Ahmed S, Sharma C, Kumar R. Hybridoma

technology a versatile method for isolation of monoclonal antibodies, its applicability across

species, limitations, advancement and future perspectives. International

immunopharmacology. 2020 Aug 1;85:106639.