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The document discusses autophagy, a controlled process of cellular self-digestion involving the formation of autophagosomes that enclose organelles and cytosol for degradation. It also briefly covers mass spectrometry techniques and their applications in analyzing biological samples, including various methods for ionization and separation of compounds. Additionally, it introduces the principle of ELISA, a technique used to detect and quantify antigens using enzyme-linked antibodies.

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20 views

C3 adds

The document discusses autophagy, a controlled process of cellular self-digestion involving the formation of autophagosomes that enclose organelles and cytosol for degradation. It also briefly covers mass spectrometry techniques and their applications in analyzing biological samples, including various methods for ionization and separation of compounds. Additionally, it introduces the principle of ELISA, a technique used to detect and quantify antigens using enzyme-linked antibodies.

Uploaded by

maittt.22ba13211
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
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Autophagy

Isolation
membrane Autophagosome

Enclosing organelles and

Yoshinori Ohsumi cytosol

Nobel Price Physiology and medicine 2016 Lysosome


Autolysosome

autophagosomes
TEM of yeast under
caloric restriction

Digestion of vesicle Digestion of internal Fusion


content membrane

vacuole
Autophagy is an auto-cannibalism complex phenomenon that is highly controled

Nocodazole
nutrient
starvation
G proteins Autophagolysosome
autophagosome
rapamycin mTOR
a.a.
Peptides
Damaged
Fatty acids
tamoxifen organites
Lipids
Sugars
Nucleotides
...
Isolation
membrane

PI3K cytochalasines Bafilomycine


Lysosome
Agrégats protéiques
Neosyntheses
Wortmannin
induction maturation Fusion/degradation SURVIVAL
Mass spectrometry in brief…

Separation techniques Mass spectrometry techniques Electronics and computation


- GC
Ion source Analysor Detector Signal processing Biostatistics
- LC en m/z
- HPLC

Abondance
- UPLC
- SDS PAGE
Under vacuum
- 2D PAGE -4 -10
or atmospheric Advanced vacuum 10 -10 mbar
- BN-PAGE pressure mass/charge (m/z)

-Nothing (shot-gun)

Ions source Analyseur Detector


- Transfer and ionise - Separate ions according m/z Ions count : convert ions
compounds in gaseous phase - Advanced vacuum container into electric signal
- Various analysers : (electrons or photons
- Various methods for soft and multiplicator)
* Time of Flight (TOF),
hard ionisation: * Quadrupole, Signal processing
soft * Ionic trap, -Graphic representation of
* Matrix Assisted Laser Desorption * Orbitrap, ions quantifications
Ionization (MALDI), * Fourier Transform Ion Cyclotron
• Electrospray (ESI), Resonance (FT-ICR)… Multivariate statistical
hard * Tandem mass Spectrometry analyses (Principal
* Electronic Ionisation (EI), (MS/MS) : provide information on componant analysis, etc …)
* Chemical Ionization (CI), structure and sequence of ions
* Fast Atom Bombardment (FAB) * Ionique mobility : isomers isobares
Various Mass spectrometry approaches

SEPARATE – SORT – ANALYSE - CLASSIFY


ELISA (enzyme-linked immunosorbent assay)
Principle
Various types of ELISA
Antigen fixed on plate
surface

Antibody with
linked enzyme
attach to antigen

Typical results of ELISA

Conc. Experimental
Scale
Substrate transformed
by enzyme into a

points
colored product
FILTER-GROWN POLARIZED CELLS

Molecules or cells can


be added apically

(plastic container)

OR molecules or cells can be


added basolaterally

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