Monoclonal Antibodies

Presented by
Tithi Roy 2008
Shams Bin Bony Amin 2009431004
Probal Sen - 2009431016
Md. Tanvir Ahmed - 2009431022

Monoclonal Antibodies
Monoclonal antibodies (MAb) are antibodies
produced from identical B immune cells.
Named Monoclonal because they come from single
type (clone) of B cells.
Recognizes a single and specific antigen.
Wide application in biochemistry, molecular biology
and meidicine.

Production method of
Monoclonal Antibodies
Production involves two phases:
Cell line production

MAb purification

Cell line production:

MAbs are produced from hybridoma cells
Hybridoma cells are fusion of antibody producing B
cell and myeloma cancer cells

Production method of
Monoclonal Antibodies
Cell line production (continued):
Mouse exposed to antigen.
Splenocytes are isolated.
B lymphocytes are collected and fused with
myeloma cells
The cell culture contain mixture of both fused and
unfused cells.

Production method of
Monoclonal Antibodies
Cell line production (continued):
Selection of Hybridoma on HAT medium
Quantitative assay of antibody by ELISA and Dot
Blotting.

Production method of
Monoclonal Antibodies

Production method of
Monoclonal Antibodies
MAb purification:
Cell debris and contaminants removed by
centrifugation.
Chromatography is useful for MAb separation.

Affinity chromatography with agarose results

high level of purification (>80%)

Production method of
Monoclonal Antibodies
MAb purification by Immunoaffinity chromatography:

Production of Recombinant
Monoclonal Antibodies
MAb produced in mice cause:
systemic inflammatory effects
Human anti-mouse antibodies (HAMA)
Solution:

mouse DNA encoding the binding portion is

replaced by human antibody-producing DNA
Produces chimeric or humanized antibodies

Applications of
Monoclonal Antibodies
In diagnostic procedures like RIA and ELISA
Hormone concentration analysis.
Diagnostic imaging using Radiolabeled MAbs.
In treatment of cancer: Detection of Tumor
specific antigen
In treatment of AIDS: Cell mediated destruction of
HIV infected cells guided by MAb

Advantages of
Monoclonal Antibodies
Production of identical antibodies at industrial
scale.
High specificity: reduced possibility of crossreaction
Better result in quantitative assay of proteins.

Disadvantages of
Monoclonal Antibodies
Technology is expensive.
Time consuming proces.

Heterogeneity of MAb often makes them

incompetent to detect target protein.
More sensitive to change of pH.
Needs particular storage facilities.

Present Applications and Future

Prospects of Nanobiotechnology

Presented by
Tithi Roy 2008
Shams Bin Bony Amin 2009431004
Probal Sen - 2009431016
Md. Tanvir Ahmed - 2009431022

From Nanotechnology
to Nanobiotechnology
Nanotechnology: The techniques of producing tiny
substances having few nanometers of size.
Exhibition of unique characteristics:
Higher volume to surface ratio
Strange optical properties: Gold
nanoparticles appear black
Decreased melting point:
Gold (slab size) = M.P. 1064C
Gold (2.5 nm size) = 300C

Nanobiotechnology
Use of Nanoparticles or nano-structured substances
on biological systems.
A relatively new discipline, rendering new promise.

Applications of Nanobiotechnology

Applications of Nanobiotechnology

In Agriculture and Food Industry

Nanofertilizers Smart fertilizers

Mode of delivery
a. Direct application of nutrient nanoparticles
b. encapsulated inside a nanoparticle
Coated outside a nanoparticle
Urea fertilized Zeolite chips slowly release nitrogen Reduced leaching and gaseous emission of Urea.
(Milan et al. 2008)

Applications of Nanobiotechnology

In Agriculture and Food Industry

Nanopesticides Nano droplets of pesticides

Higher surface area Greater contact with pests
Syngenta and BASF Developing new pesticides as
nanopartilces. (Lauterwasser et al. 2005)
Encapsulated, time-release programmed

Applications of Nanobiotechnology

In Agriculture and Food Industry

Effective detection of DNA and protein:

Employs biosensor
Useful in genetic detection of specific plant species
and plant pathogens

Quality assessment of food products:

Employs biosensor

Useful in vitamin analysis, detection of antibiotics

or microbial contamination

Applications of Nanobiotechnology

In Diagnosis and Therapeutics

Drug delivery: Surface of NPs can carry drugs,

probes or proteins.
Engineered NPs are produced from organic
compounds or organic polymers.

Potential dangers: Risk of aggregation and

cytotoxicity.

Applications of Nanobiotechnology

In Diagnosis and Therapeutics

Gene therapy: Delivery of therapeutic genes to treat

a disease
Gold NPs are good carriers to deliver the
therapeutic genes.
Biolistic method: Gene Gun
Molecular imaging: Provides detailed picture of
physiological functions.
Radioactive NPs through the bloodstream.
Special camera tracks movement.

Applications of Nanobiotechnology

In Diagnosis and Therapeutics

Biomarker mapping: NPs can detect disease

associated biomarkers at ultra-low concentrations.
Diagnosis of cancer: Cancer causing SNPs are
efficiently detected by gold NP labeled DNA
microarrays.
Abraxane - Injectable suspension for cancer
treatement

Applications of Nanobiotechnology

To conserve energy and environment

Higher efficiency in energy conversion: NPs have

enhanced absorption of solar radiation.
Good at concentrating sunlight.
Good at generating electricity in Photovoltaic cells.

Brighter light at minimum cost: Production of

energy efficient LED bulbs using nano-structure
arrays.

Applications of Nanobiotechnology

To conserve energy and environment

Enhanced productivity of windmills:

windmill blades made using carbon nanotubes.
Stronger and lighter blades
Compact and lighter fuel cell engine

Carbon nanotubes to reduce power loss in electric

transmission wires
Improved performance of rechargeable batteries

Applications of Nanobiotechnology

To conserve energy and environment

Enhanced productivity of windmills:

windmill blades made using carbon nanotubes.
Stronger and lighter blades
Compact and lighter fuel cell engine

Carbon nanotubes to reduce power loss in electric

transmission wires
Improved performance of rechargeable batteries

Future Prospects of Nanobiotechnology

Growing optimism on the advancement of
Nanobiotechnology
New concepts:

Nanodrug or nanomedicine
Targeted killing of cancer cells
Nanobots for surgery and
diagnosis
Nanogenerators