Teknik Pemeriksaan Imunologi

Pendahuluan
Verdiansah
 DEFENISI
• Imunitas
reaksi tubuh thd masuknya substansi asing
• Respon imun
kumpulan respon thd substansi asing yg
terkoordinasi
• Sistem imun
sel & molekul yg bertanggung jawad dlm
imunitas
• Imunologi
– Mekanisme fisiologi mempertahankan tubuh dari
infeksi :Bacteria,Viruses,Fungi,Parasites,Toxins
 SISTIM IMUN

1. Bawaan (the innate immune system )

•  respon imun non spesifik
2. Diperoleh (the adaptive/acquired immune
system)
•  respon imun spesifik
 Immunity
Innate immunity Adaptive immunity

Humeral Cell-mediated

Components Components
Macrophages antigen presenting cells
Granulocytes T-cells
Natural killer cells B-cells
Complement Antibodies
Other chemicals: HCL, lysozyme Complement

Characteristincs
* Action is immediate
* Response is non-specific
* Response is not enhanced on
repeated exposure to pathoge
Characteristics
* Action requires days to develop
* Response is specific
* Response is enhanced on
repeated exposure to pathogen
 Labeled immunoassays
• for antigens and antibodies that may be small
in size or present in very low concentrations.

• The presence of such antigens or antibodies is

determined indirectly by using a labeled
reactant
 Labeled immunoassays
• Radioimmunoassay

• Enzyme Immunoassay

• Rapid Immunoassays
 Membrane-Based Cassette Assays

• New type of enzyme immunoassay.

• rapid
• easy to perform
• and give reproducible results.
• semiquantitative for use in a clinical laboratory.
• single-use, disposable
 Laboratory use of antibodies
• Quantitation of an antigen
– RIA, Elisa
• Identification and characterization of protein antigens
– Immunoprecipitation
– Western blotting
• Cell surface labelling and separation
• Localisation of antigens within tissues or cells
• Expression librairies
• Phage display
 Detection principles

• Radiolabelled isotopes (antigen)

– 125I, 32P, 35S
• Enzymes (Ab)
– Peroxydase
• Chromophores (Ab)
– Fluorogenic probes (UV, visible or IR)
RIA: radio immuno assay
Typical RIA standard curve
Elisa: Enzyme-linked immunosorbent assay
Sandwich Elisa
Western blotting
ECLIA
 Flocculation test
(A precipitation reaction)

(1) Non Reactive (2) Weakly Reactive (3,4) Reactive

RPR card test

 Two dimensional electrophoresis

1st dimension 2nd dimension

Stable
Molecular weight kDa
pH gradient

pH
Immunoprecipitation

Proteomics
Western Blotting
Immunohistochemistry
 Phosphospecific antibodies to study
cellular signaling

• Phosphorylation and dephosphorylation affect

the structure and activity of proteins
• Cellular signalling is characterized by cascades
of phosphorylation
• Kinases and phosphatases maintain
phosphorylated/dephosphorylated state of
proteins
• Phospho/Tyrosine/Threonine/ Serine
Flowcytometri
R1 = limfosit
R2 = monosit
R3 = PMN
Terima kasih