O slideshow foi denunciado.
Utilizamos seu perfil e dados de atividades no LinkedIn para personalizar e exibir anúncios mais relevantes. Altere suas preferências de anúncios quando desejar.
RADIO IMMUNO ASSAY
FIRDOUS
M. Pharmacy
CONTENTS
 Introduction
 Theory
 Principle
 Requirements
 Procedure
 Applications
INTRODUCTION
• Radio immuno assay is an immunological assay to
analyse antigens present in given biological samples
• It i...
THEORY
• RIA involves the reaction between antigen (hapten) and its specific
Antibody in presence of radio labelled antige...
PRINCIPLE
• It involves three principles which make it most specific & sensitive than
other immune assays.
• An immune rea...
REQUIREMENTS
• Micro titer plate / Test tubes.
• Pure antigen
• Radio labelled antigen
• Antibodies
• Standard's
• Centrif...
Micro titer plate :
• A microtiter plate is used mostly used for this assay
• A microtiter plate could have 6, 24, 96, 384...
PURE ANTIGEN :
• Antigen may be obtained from biological sample or by synthetic form , it
should be pure.
• It is used as ...
ANTIBODY
• Specific antibodies are obtained by injecting the
Ag to animals.
• Ag I.e., drug molecule + bovineserum albumin
STANDARDS
Un labeled Ag concentration
B/F
Ratio
Centrifuge:
• Used foe the separation of
precipitated form and
supernatant liquid form.
• Range: 1200 – 2500 rpm
Radio active counters :
Two types of counters are used
They are :
a) Gamma counter.
b) Scintillation counters.
a)Gamma counter :
• These are used for the gamma energy emitting isotopes
e.g., common iodine isotopes.
b) Scintillation c...
PROCEDURE :
Test tube-1
Blank
Ab +Ag*
Test tube -2
Calibrator
Ab + pure Ag +Ag*
Test tube -3
Sample
Ab + sample Ag + Ag*
PROCEDURE :
SEPARATION TECHNIQUES :
• After completion of reaction of reaction free form and bound forms
are determined by separation ...
Applications :
Radio immuno assay of morphine :
This involves :
• Synthesis of immunogen
• Antiserum production.
• Procedu...
CONCLUSION :
• RIA is invitro assay technique used for the determination of Ag concentration
present in given sample.
• In...
REFERENCE :
RIA
RIA
RIA
Próximos SlideShares
Carregando em…5
×

RIA

Radio immuno assay is an immunological assay to
analyse antigens present in given biological samples

  • Entre para ver os comentários

RIA

  1. 1. RADIO IMMUNO ASSAY FIRDOUS M. Pharmacy
  2. 2. CONTENTS  Introduction  Theory  Principle  Requirements  Procedure  Applications
  3. 3. INTRODUCTION • Radio immuno assay is an immunological assay to analyse antigens present in given biological samples • It is most sensitive and specific method of immuno assay • Sensitivity ranges from 0.0006 -0.006 µg / ml . • It was developed by S.A. Berson and Rosalyn yalow in 1959 in received nobel prize in 1977.
  4. 4. THEORY • RIA involves the reaction between antigen (hapten) and its specific Antibody in presence of radio labelled antigen. • This hapten is obtained from a non-antigenic compound e.g., morphine , Cartelol etc., which is ultimately conjugated . • Animals normally develop antibodies to the injected immunogenic substances as part of their natural immune response. • The serum derived from this animals is used as the antibody source and tested with reference to their specificity, affinity at their titre level.
  5. 5. PRINCIPLE • It involves three principles which make it most specific & sensitive than other immune assays. • An immune reaction i.e. antigen, antibody binding. • A competitive binding or competitive displacement reaction (It gives specificity). • Measurement of radio emission (It gives sensitivity).
  6. 6. REQUIREMENTS • Micro titer plate / Test tubes. • Pure antigen • Radio labelled antigen • Antibodies • Standard's • Centrifuge • Radioactive counter
  7. 7. Micro titer plate : • A microtiter plate is used mostly used for this assay • A microtiter plate could have 6, 24, 96, 384 or even sometimes 1536 wells arranged in rows. • Each well of a microplate can only hold very small amounts of liquid.
  8. 8. PURE ANTIGEN : • Antigen may be obtained from biological sample or by synthetic form , it should be pure. • It is used as standard or calibrator. RADIO LABELLING OF ANTIGEN: •The most commonly used radiolabels in RIA are tritium and iodine • They have adequate activity and have long enough half lifes.
  9. 9. ANTIBODY • Specific antibodies are obtained by injecting the Ag to animals. • Ag I.e., drug molecule + bovineserum albumin
  10. 10. STANDARDS Un labeled Ag concentration B/F Ratio
  11. 11. Centrifuge: • Used foe the separation of precipitated form and supernatant liquid form. • Range: 1200 – 2500 rpm
  12. 12. Radio active counters : Two types of counters are used They are : a) Gamma counter. b) Scintillation counters.
  13. 13. a)Gamma counter : • These are used for the gamma energy emitting isotopes e.g., common iodine isotopes. b) Scintillation counter : • These are used for counting beta energy emitting isotopes e.g., tritium , carbon -14 isotopes
  14. 14. PROCEDURE :
  15. 15. Test tube-1 Blank Ab +Ag* Test tube -2 Calibrator Ab + pure Ag +Ag* Test tube -3 Sample Ab + sample Ag + Ag* PROCEDURE :
  16. 16. SEPARATION TECHNIQUES : • After completion of reaction of reaction free form and bound forms are determined by separation techniques. • Various techniques include gel filtration, Electrophoresis, solid phase adsorption of Ag, Ab & fractional precipitate.
  17. 17. Applications : Radio immuno assay of morphine : This involves : • Synthesis of immunogen • Antiserum production. • Procedure.
  18. 18. CONCLUSION : • RIA is invitro assay technique used for the determination of Ag concentration present in given sample. • In advance to RIA many other techniques are introduced like RAST, EIA , RIST, lRMA , competitive RIA.
  19. 19. REFERENCE :

×