Name: Slide 1
Uploaded: 2022-05-06
Duration: 49 min 30 s
Description: . Molecular Biotechnology (BIOC 408) Dr. Rania Hassan Mohamed.

. Molecular Biotechnology (BIOC 408) Dr. Rania Hassan Mohamed. 

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Molecular Biotechnology (BIOC 408)  Dr. Rania Hassan Mohamed

Therapeutic and commercial use of Biotechnology. 

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Therapeutic and commercial use of  Biotechnology

Techniques using Biotechnology: i.e. Microarray. The DNA microarray relies on cDNA fragments from a sample to hybridize with synthetic ssDNA sequences (specificA, C, G, T combinations). The synthetic fragments (oligonucleotides or oligos) are the probes. The cDNA fragments are the targets. Tat ts Probes. 

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Techniques  using Biotechnology: i.e.  Microarray

The DNA microarray
relies on cDNA fragments
from a sample to
hybridize with synthetic
ssDNA sequences
(specificA, C, G, T
combinations).
The synthetic fragments
(oligonucleotides or
oligos) are the probes.
The cDNA fragments are
the targets.
Tat ts
Probes

Techniques using Biotechnology: i.e. Microarray. DNA hybridization is when a single-stranded DNA molecule (ssDNA) reanneals with another ssDNA from another source. If the original ssDNA strand has sequences that are complementary to the introduced ssDNA strand, the two strands form a dsDNA hybrid molecule with one strand from each. Com lementar DNA bases d•DåA the rew v•.ONA to a ccrnpk•nentæy. 

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DNA hybridization is when a
single-stranded DNA
molecule (ssDNA) reanneals
with another ssDNA from
another source.
If the original ssDNA strand
has sequences that are
complementary to the
introduced ssDNA strand,
the two strands form a
dsDNA hybrid molecule with
one strand from each.
Com lementar DNA bases
d•DåA
the rew v•.ONA to a
ccrnpk•nentæy

Techniques using Biotechnology: i.e. Microarray. Each feature may contain hundreds or thousands of identical probes (oligos). Each array may contain tens of thousands of features. Each feature is looking for a specific gene sequence of nucleotide bases Thousands of specific genes can be identified simultaneously. What is the DNA sequence for the feature in the graphic?. 

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Each feature may contain
hundreds or thousands of
identical probes (oligos).
Each array may contain tens
of thousands of features.
Each feature is looking for a
specific gene sequence of
nucleotide bases
Thousands of specific genes
can be identified
simultaneously.
What is the DNA
sequence for the
feature in the
graphic?

Techniques using Biotechnology: i.e. Microarray. Cy3 (Omen) cys (Red) GONA. 

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Techniques using Biotechnology: i.e. Microarray. ///iii/'i/ö//. 

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Therapeutic use of Biotechnology : 1. Monoclonal Antibody. 

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Therapeutic use of Biotechnology :  1. Monoclonal Antibody

Variable region
Antigen-binding
site
Heavy chain
Constant region

(A)
Fab
CH3
Murine
(-momab)
(C)
Humanized
(-zumab)
(B)
Chimeric
(-ximab)
(D)
Human
(-umab)

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Antigen
of interest
Harvested B cells
Myeloma cells
Copyright Tho President and Fellows of Harvard College
Hybridomas
Hybridoma cultures
Antibodies
to antigen
of interest
Selected antigen-specific
hybridoma culture

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(A)
CHI
Human B cell mRNA
Reverse transcription to cDNA
Isotype-specific PCR for VH and VL
Cloning of PCR fragments into phagemid
vector
Electroporation into E-coli competent
helper phage rescue
Generation of phage library
(B)
Human IgG
CHI
(C)
Antigens
Phage library
generation
Immobilized antigen to
solid phase
Next panning
round
Analysis of phage
recovery titer
Fab
Phage antibody
library
Phage
amplification
scFv
Phages bind
to antigens
Wash
and
Elution
E-coli infection and
helper phage rescue

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(A) Mouse hybridoma
Immunization
with targets
Harvest
generate hybridomas
0 00
Screening
Mouse mAb
(B) Phage display
scFv or Fab
Phage-displayed Ab libraries
Biopanning
with targets
(3-5 cycles)
(C) Transgenic mouse
Immunization
with targets
Harvest splenocytes,
generate hybridomas
(D) Single B cell
Human
00
PBMC
Sort B cells with
labeled antigens
0 00
Screening
Screening
Construction of
Human IgG
Chimeric mAb
Humanized mAb
VHDJHC 1
VKJKCK
PCR, construct VH and VL
Human mAb

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Naked Antibody
Target to cancer cells,
tumor microenvironment,
immune checkpoints
Affinity maturation
Cytokine
Immunocytokine
ADCC or CDC
Point mutations or
modified glycosylation
CAR-T
T cells
Signal
domain
Antibody-Drug
Conjugate
(ADC)
Drug
Radionuclide
scFv
T cells
Target cell
Bispecific
antibody
scFv or Fab
Immunoliposome

Therapeutic use of Biotechnology : 2 . CAR-T cells. 

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Therapeutic use of Biotechnology : 2 . CAR-T cells

Single-chain variable region
Transmembrane domain
Signaling domains

First generation
Signal 1
Antigen-
recognition
domain
Signalling
domains
Second generation
Signal 2
Signal 1
Third generation
Signal 2
Signal 1
Nature Reviews I Clinical Oncology

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Make CAR T
cells in the lab
VÄVT.: •O
Insert gene
for CAR
CAR T-Cell Therapy
T cell
protein
1
O
Crow millions
of CAR T cells
CAR T cell
Infuse
o
Remove from
patient to get T cells
Cancer cell
o
CAR T cells
into patient
CAR T cells bind
to cancer cells
and kill them
Antigen

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Antigen Escape
CAR-T Cells
Cancer Cell with Cancer Cell without
Target Antigen Target Antigen
C• CAR-T cell Trafficking and D•
Infiltration
MDSC
Cancer
Cells
On Target Off Tumor
CAR T cells
Cancer Cell with Normal Cell with
Target Antigen Target Antigen
Immunosuppressive
Microenvironment
Treg
MDSC
O
Fibroblasts
CAR-T Cell
CAR.
Cell
o
Fibroblasts

E.
CAR-T Cell Associated
Toxicities
CAR-T cell
Cancer Cell
Myeloid Cell

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Table displaying FDA approved CAR T-Cell therapies.

Therapeutic use of Biotechnology : More commercial products 1- Antibiotics. 

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Therapeutic use of Biotechnology : More commercial  products 1- Antibiotics

I. Isolation
Spread a soil
dilution on a plate
of selective
medium
Sterile glass
spreader
Incubation
Colonies of
Streptomyces
species

Overlay With an
indicator organism
Incubate
Nonproducing
organisms
Zones of
growth
inhibition
Producing
organisms

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II. Testing Activity Spectrum
Streak antibiotic
producer across
one side of plate
Incubate to permit
growth and antibiotic
production
Antibiotic
diffuses
into agar
Streptomyces
cell mass

Cross-streak with
test organisms
Incubate to permit
test organisms to grow
Growth of
test organism
Inhibition zones
where sensitive
test organisms
did not grow

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Table 15-3 Some antibiotics produced commerciallya
Antibiotic
Bacitracin
Cephalosporin
Cycloheximide
Cycloserine
Erythromycin
Griseofulvin
Kanamycin
Lincomycin
Neomycin
Nystatin
penicillin
Polymyxin B
Streptomycin
Tetracycline
Producing microorganism
Bacillus licheniformis (EFB)
Cephalosporium spp. (F)
Streptomyces griseus (A)
Streptomyces orchidaceus (A)
Streptomyces erythreus (A)
Penicillium griseofulvum (F)
Streptomyces kanamyceticus (A)
Streptomyces lincolnensis (A)
Streptomyces fradiae (A)
Streptomyces noursei (A)
Penicillium chrysogenum (F)
Bacillus polymyxa (EFB)
Streptomyces griseus (A)
Streptomyces rimosus (A)
asee Chapter 26 for structures and more discussion of these antibiotics.
bEFB, endospore-forming bacterium; F, fungus; A, actinomycete.

Therapeutic use of Biotechnology : More commercial products 2 - Enzymes. 

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Therapeutic use of Biotechnology : More commercial  products 2 - Enzymes

Carrier-bound
enzyme
Enzyme inclusion in
fibrous polymers
Cross-linked
enzyme
eye
Enzyme inclusion
in microcapsules

(u) ew!1
+80 snid 000LL
00011 —
00001 —
0006 —
seppeqooes06!104—
eseuelnllnd
00 L

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