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IMSE (FINALS) > precipitation > Flashcards

precipitation Flashcards

1
Q

electro-immunodiffusion

  • When RID is applied with current, it becomes electroimmunoassay/ rocket electrophoresis
  • Ag and Ab different charges at selected pH
  • Band coverage finally meet at the middle, when all antigen is precipitated , the precipitin pattern resembles that of a shooting rocket.
A

ROCKET ELECTROPHORESIS/ LAURELL TECHNIQUE
(ONE DIMENSIONAL SINGLE ELECTROPHORESIS)

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2
Q

immunodiffusion

Ag and Ab diffuse radially

A

double dimension

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3
Q

passive immunodiffusion

  • immunoglobulins, complement
  • Ab in gel + Ag in well&raquo_space; diffusion
  • precipitin ring
A

radial immunodiffusion (RID)

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4
Q

patterns

crossed bands

A

non-identity

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5
Q

electro-immunodiffusion

  • Antigen and antibody migrate toward each other by electrophoresis.
  • Used only when Ag and Ab have opposite charges.
  • Detection of the semi-quantitative antigen of the body fluid
  • Faster and more sensitive than Ouchterlony Method
A

COUNTERCURRENT IMMUNOELETROPHORESIS (ONE DIMENSIONAL-DOUBLE ELECTROPHORESIS)

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6
Q

The strength with which a multivalent antibody binds a multivalent antigen.

A

avidity

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7
Q
  • A semiquantitative gel precipitation technique in which proteins are first separated by electrophoresis and then subjected to double diffusion with antibodies directed against the individual proteins.
  • Reliable and accurate method fro detecting structural abnormalities and concentration changes in proteins
  • Useful in screening circulating immune complexes/ Myeloma proteins
  • METHOD
    1) Ag is separated by electrophoresis,
    2) Ab is placed in trough cut in the agar,
    3) Trough is filled with an anti-serum
    4) Incubation: 18 to 24 hours
  • APPLICATIONS
    1) Serum: detection of Monoclonal Gammopathy
    2) Urine: detection of Bence Jones Protein
A

immuno electrophoresis

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8
Q
  • Immunodiffusion reaction in a support medium with the use of electric current to enhance mobility of reactants and to increase movement toward one another.
  • PRINCIPLE:
    1) Antibody: Positively Charged: Migrates toward the cathode (-)
    2) Antigen: Negatively Charged: Migrates toward the anode (+)
A

electro-immunodiffusion

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9
Q
  • ELECTRO-IMMUNODIFFUSION
  • IMMUNO-ELECTROPHORESIS
  • IMMUNO-FIXATION
A

electrophoretic method

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10
Q

types

  • RADIAL IMMUNODIFFUSION
  • OUCHTERLONY DOUBLE DIFFUSION
A

passive immunodiffusion method

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11
Q
  • Soluble Antigen (Kraus, 1897).
  • The combination of soluble antigen with soluble antibody to produce visible insoluble complexes.
  • SOLUBLE ANTIGEN + SOLUBLE ANTIBODY → INSOLUBLE COMPLEX
A

precipitation

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12
Q

passive immunodiffusion

  • 27-72 hrs
  • IgG: 24 hrs
  • IgM: 50-72 hrs
  • SQUARE of diameter is directly proportional to concentration
  • precipitin ring
A

mancini (endpoint)

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13
Q

Antibodies are capable of reacting with antigens that are structurally similar to the original antigen that induced antibody production

A

cross-reactivity

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14
Q

immunodiffusion

both Ag and Ab are moving

A

double diffusion

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15
Q

passive immunodiffusion

  • precipitation (rabbit)
  • flocculation (horse)
  • Ab in agarose + Ag on top&raquo_space; Ag diffuses downward
  • precipitation occurs in proportion to the amount of Ag present
A

OUDIN

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16
Q

radial immunodiffusion

  • Antigen is allowed to diffuse to completionand when equivalence is reached, there is no further change in the ring diameter
  • longer reaction time (sensitive)
  • occur 24 - 72 hours
  • ring diameter = antigen conc.
  • uses graphing paper
A

mancini (endpoint diffusion)

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17
Q

patterns

  • fusion of two lines with a spur
  • share a common epitope but some Ab are not captured by Ag
A

partial identity

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18
Q

immunodiffusion

pertains to the antigen or antibody

A

“immuno”

19
Q

precipitation

  • VDRL and other non-treponemal tests (except: RPR)
  • standardization of toxins and toxoids
  • a specific type of precipitation that occurs over a narrow range of antigen concentrations
  • antigen consists of very fine particles that Ag-Ab complexes remain suspended instead of settling as sediments
  • may require microscopy
A

flocculation

20
Q
A
21
Q

combination

double diffusion-single dimension

A

Oakley and Fulthrope

22
Q

patterns

fused bands

A

serological identity

23
Q

immunodiffusion

only one reactant is moving (usually Ag)

A

single diffusion

24
Q

types

  • TURBIDIMETRY
  • NEPHELOMETRY
A

precipitation by light scatter

25
Q

passive immunodiffusion

  • complex antigens
  • fungal antigens
  • both Ag and Ab diffuse in two dimensions
  • PARTIAL ID = diff. Ags but similar epitopes
  • precipitin paterns: X (non identity), C (identity), K (partial ID)
A

ouchterlony

26
Q

A law used to mathematically describe the equilibrium relationship between soluble reactants and insoluble products. It can be applied to antigen–antibody relationships

A

law of mass action

27
Q

immunodiffusion

movement of the antigen or antibody

A

“diffusion”

28
Q

combination

single-diffusion-double dimension

A

Radial Immunodiffusion/ Mancini

29
Q
  • A simple , specific method for identification and quantification of a number of proteins found in serum and other body fluids.
  • Internal reactants like specific antibodies added to buffered agarose medium, and serum containing standard volume of CHON or Ag is placed in well, centered in agarose.
  • Result: Precipitin Ring
  • Interpretation: Diameter of the ring is proportional to the concentration of the Antigen.
  • Quantitative: Ig Levels, Serum CHON’s, Serum Complement
A

radial immunodiffusion

30
Q

combination

double-diffusion-double dimension

A

Ouchterlony and Elek

31
Q

radial immunodiffusion

  • Measurement taken before the point of equivalence is reached, Antigen isnot allowed to diffuse completely
  • shorter reaction time
  • read wihtin 18 hours (24 hours)
  • ring diameter = log conc. (Ag)
  • uses semi-log paper
A

fahey & Mckelvey (kinetic diffusion)

32
Q

precipitation

  • lancefield
  • neufeld quellung
  • carbohydrate cell wall Ag
  • capsular swelling Ag
A

liquid precipitation

33
Q
  • A detection device is placed in direct line with the incident light, collecting light after it has passed through the solution.
  • Measures the reduction in light intensity due to reflection, absorption, or scatter.
  • Scattering of light occurs in proportion to the size, shape, and concentration of molecules present in solution.
A

turbidimetry

34
Q
  • Ag/Ab Complexes form and precipitate when the 2 reactants meet at the equivalence zone.
  • Use to determine the relationship between Ag and Ab
  • Both Ag and Ab diffuse
  • Clinical application: Detection of antibody associated with autoimmune disease (RA, SLE Sjogren’s)
  • Disadvantage: Semiquantitative only
A

ouchterlony double diffusion

35
Q

combination

Single diffusion- single dimension

A

OUDIN

36
Q
  • Measures the light that is scattered at a particular angle from the incident beam as it passes through a suspension.
  • measure light scatter at angles ranging from 10 degrees to about 90 degrees.
  • Light scatter may be recorded in arbitrary units of “relative light scatter,” or it may be directly extrapolated by a computer to give actual concentrations in milligrams per deciliter (mg/dL) or international units per milliliter (IU/mL)
A

nephelometry

37
Q

passive immunodiffusion

plain gel is placed between Ag and Ab to fascilitate double diffusion

A

Oakley And Fulthrope

38
Q

Is the initial force of attraction that exists between a site on an antibody molecule and a single epitope single Fab or determinant site on the corresponding antigen.

A

affinity

39
Q

immunodiffusion

the movement are either UP or DOWN

A

single dimension

40
Q

passive immunodiffusion

  • 18-19 hours (24hrs)
  • diameter is proportional to the LOG of the concentration
  • precipitin ring
A

fahey and Mckelvy (kinetic)

41
Q

gel precipitation

  • Both reactants (Ag and Ab) diffuse within a gel.
  • Both reagents are addedafter gel has set.
A

DOUBLE IMMUNODIFFUSION TECHNIQUE

42
Q

gel precipitation

  • One reactant (Ag or Ab) remains fixed in gel.
  • Other reactant is allowed to moved.
  • Interaction with the reagent that is immobilized.
  • Reagents become fixed in the gel if they are added to the gel medium while it is in liquid form.
A

SINGLE IMMUNODIFFUSION TECHNIQUE

43
Q
  • A semiquantitative gel precipitation technique similar to that of immunoelectrophoresis, except that antibody is added directly to the surface of the gel after electrophoresis has taken place.
  • A cellulose acetate strip impregnated with antiserum is placed over the separate proteins after serum , urine or CSF are electrophoresed.
  • Diffusion of antiserum into the gel occurs rapidly, resulting in precipitation
A

immuno-fixation electrophoresis

IMSE (FINALS) (8 decks)

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